sample purpose statement for qualitative research

From John W. Creswell \(2016\). 30 Essential Skills for the Qualitative Researcher \ . Thousand Oaks, CA: Sage.

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Purpose Statement Overview

Best practices for writing your purpose statement, writing your purpose statement, sample purpose statements.

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The purpose statement succinctly explains (on no more than 1 page) the objectives of the research study. These objectives must directly address the problem and help close the stated gap. Expressed as a formula:

Good purpose statements:

• Flow from the problem statement and actually address the proposed problem
• Are concise and clear
• Answer the question ‘Why are you doing this research?’
• Match the methodology (similar to research questions)
• Have a ‘hook’ to get the reader’s attention
• Set the stage by clearly stating, “The purpose of this (qualitative or quantitative) study is to ...

In PhD studies, the purpose usually involves applying a theory to solve the problem. In other words, the purpose tells the reader what the goal of the study is, and what your study will accomplish, through which theoretical lens. The purpose statement also includes brief information about direction, scope, and where the data will come from.

A problem and gap in combination can lead to different research objectives, and hence, different purpose statements. In the example from above where the problem was severe underrepresentation of female CEOs in Fortune 500 companies and the identified gap related to lack of research of male-dominated boards; one purpose might be to explore implicit biases in male-dominated boards through the lens of feminist theory. Another purpose may be to determine how board members rated female and male candidates on scales of competency, professionalism, and experience to predict which candidate will be selected for the CEO position. The first purpose may involve a qualitative ethnographic study in which the researcher observes board meetings and hiring interviews; the second may involve a quantitative regression analysis. The outcomes will be very different, so it’s important that you find out exactly how you want to address a problem and help close a gap!

The purpose of the study must not only align with the problem and address a gap; it must also align with the chosen research method. In fact, the DP/DM template requires you to name the  research method at the very beginning of the purpose statement. The research verb must match the chosen method. In general, quantitative studies involve “closed-ended” research verbs such as determine , measure , correlate , explain , compare , validate , identify , or examine ; whereas qualitative studies involve “open-ended” research verbs such as explore , understand , narrate , articulate [meanings], discover , or develop .

A qualitative purpose statement following the color-coded problem statement (assumed here to be low well-being among financial sector employees) + gap (lack of research on followers of mid-level managers), might start like this:

In response to declining levels of employee well-being, the purpose of the qualitative phenomenology was to explore and understand the lived experiences related to the well-being of the followers of novice mid-level managers in the financial services industry. The levels of follower well-being have been shown to correlate to employee morale, turnover intention, and customer orientation (Eren et al., 2013). A combined framework of Leader-Member Exchange (LMX) Theory and the employee well-being concept informed the research questions and supported the inquiry, analysis, and interpretation of the experiences of followers of novice managers in the financial services industry.

A quantitative purpose statement for the same problem and gap might start like this:

In response to declining levels of employee well-being, the purpose of the quantitative correlational study was to determine which leadership factors predict employee well-being of the followers of novice mid-level managers in the financial services industry. Leadership factors were measured by the Leader-Member Exchange (LMX) assessment framework  by Mantlekow (2015), and employee well-being was conceptualized as a compound variable consisting of self-reported turnover-intent and psychological test scores from the Mental Health Survey (MHS) developed by Johns Hopkins University researchers.

Both of these purpose statements reflect viable research strategies and both align with the problem and gap so it’s up to the researcher to design a study in a manner that reflects personal preferences and desired study outcomes. Note that the quantitative research purpose incorporates operationalized concepts  or variables ; that reflect the way the researcher intends to measure the key concepts under study; whereas the qualitative purpose statement isn’t about translating the concepts under study as variables but instead aim to explore and understand the core research phenomenon.  

Always keep in mind that the dissertation process is iterative, and your writing, over time, will be refined as clarity is gradually achieved. Most of the time, greater clarity for the purpose statement and other components of the Dissertation is the result of a growing understanding of the literature in the field. As you increasingly master the literature you will also increasingly clarify the purpose of your study.

The purpose statement should flow directly from the problem statement. There should be clear and obvious alignment between the two and that alignment will get tighter and more pronounced as your work progresses.

The purpose statement should specifically address the reason for conducting the study, with emphasis on the word specifically. There should not be any doubt in your readers’ minds as to the purpose of your study. To achieve this level of clarity you will need to also insure there is no doubt in your mind as to the purpose of your study.

Many researchers benefit from stopping your work during the research process when insight strikes you and write about it while it is still fresh in your mind. This can help you clarify all aspects of a dissertation, including clarifying its purpose.

Your Chair and your committee members can help you to clarify your study’s purpose so carefully attend to any feedback they offer.

The purpose statement should reflect the research questions and vice versa. The chain of alignment that began with the research problem description and continues on to the research purpose, research questions, and methodology must be respected at all times during dissertation development. You are to succinctly describe the overarching goal of the study that reflects the research questions. Each research question narrows and focuses the purpose statement. Conversely, the purpose statement encompasses all of the research questions.

Identify in the purpose statement the research method as quantitative, qualitative or mixed (i.e., “The purpose of this [qualitative/quantitative/mixed] study is to ...)

Avoid the use of the phrase “research study” since the two words together are redundant.

Follow the initial declaration of purpose with a brief overview of how, with what instruments/data, with whom and where (as applicable) the study will be conducted. Identify variables/constructs and/or phenomenon/concept/idea. Since this section is to be a concise paragraph, emphasis must be placed on the word brief. However, adding these details will give your readers a very clear picture of the purpose of your research.

Developing the purpose section of your dissertation is usually not achieved in a single flash of insight. The process involves a great deal of reading to find out what other scholars have done to address the research topic and problem you have identified. The purpose section of your dissertation could well be the most important paragraph you write during your academic career, and every word should be carefully selected. Think of it as the DNA of your dissertation. Everything else you write should emerge directly and clearly from your purpose statement. In turn, your purpose statement should emerge directly and clearly from your research problem description. It is good practice to print out your problem statement and purpose statement and keep them in front of you as you work on each part of your dissertation in order to insure alignment.

It is helpful to collect several dissertations similar to the one you envision creating. Extract the problem descriptions and purpose statements of other dissertation authors and compare them in order to sharpen your thinking about your own work.  Comparing how other dissertation authors have handled the many challenges you are facing can be an invaluable exercise. Keep in mind that individual universities use their own tailored protocols for presenting key components of the dissertation so your review of these purpose statements should focus on content rather than form.

Once your purpose statement is set it must be consistently presented throughout the dissertation. This may require some recursive editing because the way you articulate your purpose may evolve as you work on various aspects of your dissertation. Whenever you make an adjustment to your purpose statement you should carefully follow up on the editing and conceptual ramifications throughout the entire document.

In establishing your purpose you should NOT advocate for a particular outcome. Research should be done to answer questions not prove a point. As a researcher, you are to inquire with an open mind, and even when you come to the work with clear assumptions, your job is to prove the validity of the conclusions reached. For example, you would not say the purpose of your research project is to demonstrate that there is a relationship between two variables. Such a statement presupposes you know the answer before your research is conducted and promotes or supports (advocates on behalf of) a particular outcome. A more appropriate purpose statement would be to examine or explore the relationship between two variables.

Your purpose statement should not imply that you are going to prove something. You may be surprised to learn that we cannot prove anything in scholarly research for two reasons. First, in quantitative analyses, statistical tests calculate the probability that something is true rather than establishing it as true. Second, in qualitative research, the study can only purport to describe what is occurring from the perspective of the participants. Whether or not the phenomenon they are describing is true in a larger context is not knowable. We cannot observe the phenomenon in all settings and in all circumstances.

It is important to distinguish in your mind the differences between the Problem Statement and Purpose Statement.

The Problem Statement is why I am doing the research

The Purpose Statement is what type of research I am doing to fit or address the problem

The Purpose Statement includes:

• Method of Study
• Specific Population

Remember, as you are contemplating what to include in your purpose statement and then when you are writing it, the purpose statement is a concise paragraph that describes the intent of the study, and it should flow directly from the problem statement.  It should specifically address the reason for conducting the study, and reflect the research questions.  Further, it should identify the research method as qualitative, quantitative, or mixed.  Then provide a brief overview of how the study will be conducted, with what instruments/data collection methods, and with whom (subjects) and where (as applicable). Finally, you should identify variables/constructs and/or phenomenon/concept/idea.

Qualitative Purpose Statement

Creswell (2002) suggested for writing purpose statements in qualitative research include using deliberate phrasing to alert the reader to the purpose statement. Verbs that indicate what will take place in the research and the use of non-directional language that do not suggest an outcome are key. A purpose statement should focus on a single idea or concept, with a broad definition of the idea or concept. How the concept was investigated should also be included, as well as participants in the study and locations for the research to give the reader a sense of with whom and where the study took place. 

Creswell (2003) advised the following script for purpose statements in qualitative research:

“The purpose of this qualitative_________________ (strategy of inquiry, such as ethnography, case study, or other type) study is (was? will be?) to ________________ (understand? describe? develop? discover?) the _________________(central phenomenon being studied) for ______________ (the participants, such as the individual, groups, organization) at __________(research site). At this stage in the research, the __________ (central phenomenon being studied) will be generally defined as ___________________ (provide a general definition)” (pg. 90).

Quantitative Purpose Statement

Creswell (2003) offers vast differences between the purpose statements written for qualitative research and those written for quantitative research, particularly with respect to language and the inclusion of variables. The comparison of variables is often a focus of quantitative research, with the variables distinguishable by either the temporal order or how they are measured. As with qualitative research purpose statements, Creswell (2003) recommends the use of deliberate language to alert the reader to the purpose of the study, but quantitative purpose statements also include the theory or conceptual framework guiding the study and the variables that are being studied and how they are related. 

Creswell (2003) suggests the following script for drafting purpose statements in quantitative research:

“The purpose of this _____________________ (experiment? survey?) study is (was? will be?) to test the theory of _________________that _________________ (compares? relates?) the ___________(independent variable) to _________________________(dependent variable), controlling for _______________________ (control variables) for ___________________ (participants) at _________________________ (the research site). The independent variable(s) _____________________ will be generally defined as _______________________ (provide a general definition). The dependent variable(s) will be generally defined as _____________________ (provide a general definition), and the control and intervening variables(s), _________________ (identify the control and intervening variables) will be statistically controlled in this study” (pg. 97).

• The purpose of this qualitative study was to determine how participation in service-learning in an alternative school impacted students academically, civically, and personally.  There is ample evidence demonstrating the failure of schools for students at-risk; however, there is still a need to demonstrate why these students are successful in non-traditional educational programs like the service-learning model used at TDS.  This study was unique in that it examined one alternative school’s approach to service-learning in a setting where students not only serve, but faculty serve as volunteer teachers.  The use of a constructivist approach in service-learning in an alternative school setting was examined in an effort to determine whether service-learning participation contributes positively to academic, personal, and civic gain for students, and to examine student and teacher views regarding the overall outcomes of service-learning.  This study was completed using an ethnographic approach that included observations, content analysis, and interviews with teachers at The David School.
• The purpose of this quantitative non-experimental cross-sectional linear multiple regression design was to investigate the relationship among early childhood teachers’ self-reported assessment of multicultural awareness as measured by responses from the Teacher Multicultural Attitude Survey (TMAS) and supervisors’ observed assessment of teachers’ multicultural competency skills as measured by the Multicultural Teaching Competency Scale (MTCS) survey. Demographic data such as number of multicultural training hours, years teaching in Dubai, curriculum program at current school, and age were also examined and their relationship to multicultural teaching competency. The study took place in the emirate of Dubai where there were 14,333 expatriate teachers employed in private schools (KHDA, 2013b).
• The purpose of this quantitative, non-experimental study is to examine the degree to which stages of change, gender, acculturation level and trauma types predicts the reluctance of Arab refugees, aged 18 and over, in the Dearborn, MI area, to seek professional help for their mental health needs. This study will utilize four instruments to measure these variables: University of Rhode Island Change Assessment (URICA: DiClemente & Hughes, 1990); Cumulative Trauma Scale (Kira, 2012); Acculturation Rating Scale for Arabic Americans-II Arabic and English (ARSAA-IIA, ARSAA-IIE: Jadalla & Lee, 2013), and a demographic survey. This study will examine 1) the relationship between stages of change, gender, acculturation levels, and trauma types and Arab refugees’ help-seeking behavior, 2) the degree to which any of these variables can predict Arab refugee help-seeking behavior.  Additionally, the outcome of this study could provide researchers and clinicians with a stage-based model, TTM, for measuring Arab refugees’ help-seeking behavior and lay a foundation for how TTM can help target the clinical needs of Arab refugees. Lastly, this attempt to apply the TTM model to Arab refugees’ condition could lay the foundation for future research to investigate the application of TTM to clinical work among refugee populations.
• The purpose of this qualitative, phenomenological study is to describe the lived experiences of LLM for 10 EFL learners in rural Guatemala and to utilize that data to determine how it conforms to, or possibly challenges, current theoretical conceptions of LLM. In accordance with Morse’s (1994) suggestion that a phenomenological study should utilize at least six participants, this study utilized semi-structured interviews with 10 EFL learners to explore why and how they have experienced the motivation to learn English throughout their lives. The methodology of horizontalization was used to break the interview protocols into individual units of meaning before analyzing these units to extract the overarching themes (Moustakas, 1994). These themes were then interpreted into a detailed description of LLM as experienced by EFL students in this context. Finally, the resulting description was analyzed to discover how these learners’ lived experiences with LLM conformed with and/or diverged from current theories of LLM.
• The purpose of this qualitative, embedded, multiple case study was to examine how both parent-child attachment relationships are impacted by the quality of the paternal and maternal caregiver-child interactions that occur throughout a maternal deployment, within the context of dual-military couples. In order to examine this phenomenon, an embedded, multiple case study was conducted, utilizing an attachment systems metatheory perspective. The study included four dual-military couples who experienced a maternal deployment to Operation Iraqi Freedom (OIF) or Operation Enduring Freedom (OEF) when they had at least one child between 8 weeks-old to 5 years-old.  Each member of the couple participated in an individual, semi-structured interview with the researcher and completed the Parenting Relationship Questionnaire (PRQ). “The PRQ is designed to capture a parent’s perspective on the parent-child relationship” (Pearson, 2012, para. 1) and was used within the proposed study for this purpose. The PRQ was utilized to triangulate the data (Bekhet & Zauszniewski, 2012) as well as to provide some additional information on the parents’ perspective of the quality of the parent-child attachment relationship in regards to communication, discipline, parenting confidence, relationship satisfaction, and time spent together (Pearson, 2012). The researcher utilized the semi-structured interview to collect information regarding the parents' perspectives of the quality of their parental caregiver behaviors during the deployment cycle, the mother's parent-child interactions while deployed, the behavior of the child or children at time of reunification, and the strategies or behaviors the parents believe may have contributed to their child's behavior at the time of reunification. The results of this study may be utilized by the military, and by civilian providers, to develop proactive and preventive measures that both providers and parents can implement, to address any potential adverse effects on the parent-child attachment relationship, identified through the proposed study. The results of this study may also be utilized to further refine and understand the integration of attachment theory and systems theory, in both clinical and research settings, within the field of marriage and family therapy.

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9 Examples: How to Write a Purpose Statement

• Key Elements of a Purpose Statement Part 1
• How to Write a Purpose Statement Step-by-Step Part 2
• Identifying Your Goals Part 3
• Defining Your Audience Part 4
• Outlining Your Methods Part 5
• Stating the Expected Outcomes Part 6
• Purpose Statement Example for a Research Paper Part 7
• Purpose Statement Example For Personal Goals Part 8
• Purpose Statement Example For Business Objectives Part 9
• Purpose Statement Example For an Essay Part 10
• Purpose Statement Example For a Proposal Part 11
• Purpose Statement Example For a Report Part 12
• Purpose Statement Example For a Project Part 13
• Purpose Statement Templates Part 14

A purpose statement is a vital component of any project, as it sets the tone for the entire piece of work. It tells the reader what the project is about, why it’s important, and what the writer hopes to achieve.

Part 1 Key Elements of a Purpose Statement

When writing a purpose statement, there are several key elements that you should keep in mind. These elements will help you to create a clear, concise, and effective statement that accurately reflects your goals and objectives.

1. The Problem or Opportunity

The first element of a purpose statement is the problem or opportunity that you are addressing. This should be a clear and specific description of the issue that you are trying to solve or the opportunity that you are pursuing.

2. The Target Audience

The second element is the target audience for your purpose statement. This should be a clear and specific description of the group of people who will benefit from your work.

3. The Solution

The third element is the solution that you are proposing. This should be a clear and specific description of the action that you will take to address the problem or pursue the opportunity.

4. The Benefits

The fourth element is the benefits that your solution will provide. This should be a clear and specific description of the positive outcomes that your work will achieve.

5. The Action Plan

The fifth element is the action plan that you will follow to implement your solution. This should be a clear and specific description of the steps that you will take to achieve your goals.

Part 2 How to Write a Purpose Statement Step-by-Step

Writing a purpose statement is an essential part of any research project. It helps to clarify the purpose of your study and provides direction for your research. Here are some steps to follow when writing a purpose statement:

• Start with a clear research question: The first step in writing a purpose statement is to have a clear research question. This question should be specific and focused on the topic you want to research.
• Identify the scope of your study: Once you have a clear research question, you need to identify the scope of your study. This involves determining what you will and will not include in your research.
• Define your research objectives: Your research objectives should be specific, measurable, achievable, relevant, and time-bound. They should also be aligned with your research question and the scope of your study.
• Determine your research design: Your research design will depend on the nature of your research question and the scope of your study. You may choose to use a qualitative, quantitative, or mixed-methods approach.
• Write your purpose statement: Your purpose statement should be a clear and concise statement that summarizes the purpose of your study. It should include your research question, the scope of your study, your research objectives, and your research design.

Research question: What are the effects of social media on teenage mental health?

Scope of study: This study will focus on teenagers aged 13-18 in the United States.

Research objectives: To determine the prevalence of social media use among teenagers, to identify the types of social media used by teenagers, to explore the relationship between social media use and mental health, and to provide recommendations for parents, educators, and mental health professionals.

Research design: This study will use a mixed-methods approach, including a survey and interviews with teenagers and mental health professionals.

Purpose statement: The purpose of this study is to examine the effects of social media on teenage mental health among teenagers aged 13-18 in the United States. The study will use a mixed-methods approach, including a survey and interviews with teenagers and mental health professionals. The research objectives are to determine the prevalence of social media use among teenagers, to identify the types of social media used by teenagers, to explore the relationship between social media use and mental health, and to provide recommendations for parents, educators, and mental health professionals.

Part 3 Section 1: Identifying Your Goals

Before you start writing your purpose statement, it’s important to identify your goals. To do this, ask yourself the following questions:

• What do I want to achieve?
• What problem do I want to solve?
• What impact do I want to make?

Once you have a clear idea of your goals, you can start crafting your purpose statement. Your purpose statement should be a clear and concise statement that outlines the purpose of your work.

For example, if you’re writing a purpose statement for a business, your statement might look something like this:

“Our purpose is to provide high-quality products and services that improve the lives of our customers and contribute to the growth and success of our company.”

If you’re writing a purpose statement for a non-profit organization, your statement might look something like this:

“Our purpose is to improve the lives of underserved communities by providing access to education, healthcare, and other essential services.”

Remember, your purpose statement should be specific, measurable, and achievable. It should also be aligned with your values and goals, and it should inspire and motivate you to take action.

Part 4 Section 2: Defining Your Audience

Once you have established the purpose of your statement, it’s important to consider who your audience is. The audience for your purpose statement will depend on the context in which it will be used. For example, if you’re writing a purpose statement for a research paper, your audience will likely be your professor or academic peers. If you’re writing a purpose statement for a business proposal, your audience may be potential investors or clients.

Defining your audience is important because it will help you tailor your purpose statement to the specific needs and interests of your readers. You want to make sure that your statement is clear, concise, and relevant to your audience.

To define your audience, consider the following questions:

• Who will be reading your purpose statement?
• What is their level of knowledge or expertise on the topic?
• What are their needs and interests?
• What do they hope to gain from reading your purpose statement?

Once you have a clear understanding of your audience, you can begin to craft your purpose statement with their needs and interests in mind. This will help ensure that your statement is effective in communicating your goals and objectives to your readers.

For example, if you’re writing a purpose statement for a research paper on the effects of climate change on agriculture, your audience may be fellow researchers in the field of environmental science. In this case, you would want to make sure that your purpose statement is written in a way that is clear and concise, using technical language that is familiar to your audience.

Or, if you’re writing a purpose statement for a business proposal to potential investors, your audience may be less familiar with the technical aspects of your project. In this case, you would want to make sure that your purpose statement is written in a way that is easy to understand, using clear and concise language that highlights the benefits of your proposal.

The key to defining your audience is to put yourself in their shoes and consider what they need and want from your purpose statement.

Part 5 Section 3: Outlining Your Methods

After you have identified the purpose of your statement, it is time to outline your methods. This section should describe how you plan to achieve your goal and the steps you will take to get there. Here are a few tips to help you outline your methods effectively:

• Start with a general overview: Begin by providing a brief overview of the methods you plan to use. This will give your readers a sense of what to expect in the following paragraphs.
• Break down your methods: Break your methods down into smaller, more manageable steps. This will make it easier for you to stay organized and for your readers to follow along.
• Use bullet points: Bullet points can help you organize your ideas and make your methods easier to read. Use them to list the steps you will take to achieve your goal.
• Be specific: Make sure you are specific about the methods you plan to use. This will help your readers understand exactly what you are doing and why.
• Provide examples: Use examples to illustrate your methods. This will make it easier for your readers to understand what you are trying to accomplish.

Part 6 Section 4: Stating the Expected Outcomes

After defining the problem and the purpose of your research, it’s time to state the expected outcomes. This is where you describe what you hope to achieve by conducting your research. The expected outcomes should be specific and measurable, so you can determine if you have achieved your goals.

It’s important to be realistic when stating your expected outcomes. Don’t make exaggerated or false claims, and don’t promise something that you can’t deliver. Your expected outcomes should be based on your research question and the purpose of your study.

Here are some examples of expected outcomes:

• To identify the factors that contribute to employee turnover in the company.
• To develop a new marketing strategy that will increase sales by 20% within the next year.
• To evaluate the effectiveness of a new training program for improving customer service.
• To determine the impact of social media on consumer behavior.

When stating your expected outcomes, make sure they align with your research question and purpose statement. This will help you stay focused on your goals and ensure that your research is relevant and meaningful.

In addition to stating your expected outcomes, you should also describe how you will measure them. This could involve collecting data through surveys, interviews, or experiments, or analyzing existing data from sources such as government reports or industry publications.

Part 7 Purpose Statement Example for a Research Paper

If you are writing a research paper, your purpose statement should clearly state the objective of your study. Here is an example of a purpose statement for a research paper:

The purpose of this study is to investigate the effects of social media on the mental health of teenagers in the United States.

This purpose statement clearly states the objective of the study and provides a specific focus for the research.

Part 8 Purpose Statement Example For Personal Goals

When writing a purpose statement for your personal goals, it’s important to clearly define what you want to achieve and why. Here’s a template that can help you get started:

“I want to [goal] so that [reason]. I will achieve this by [action].”

Example: “I want to lose 10 pounds so that I can feel more confident in my body. I will achieve this by going to the gym three times a week and cutting out sugary snacks.”

Remember to be specific and realistic when setting your goals and actions, and to regularly review and adjust your purpose statement as needed.

Part 9 Purpose Statement Example For Business Objectives

If you’re writing a purpose statement for a business objective, this template can help you get started:

[Objective] [Action verb] [Target audience] [Outcome or benefit]

Here’s an example using this template:

Increase online sales by creating a more user-friendly website for millennial shoppers.

This purpose statement is clear and concise. It identifies the objective (increase online sales), the action verb (creating), the target audience (millennial shoppers), and the outcome or benefit (a more user-friendly website).

Part 10 Purpose Statement Example For an Essay

“The purpose of this essay is to examine the causes and consequences of climate change, with a focus on the role of human activities, and to propose solutions that can mitigate its impact on the environment and future generations.”

This purpose statement clearly states the subject of the essay (climate change), what aspects will be explored (causes, consequences, human activities), and the intended outcome (proposing solutions). It provides a clear roadmap for the reader and sets the direction for the essay.

Part 11 Purpose Statement Example For a Proposal

“The purpose of this proposal is to secure funding and support for the establishment of a community garden in [Location], aimed at promoting sustainable urban agriculture, fostering community engagement, and improving local access to fresh, healthy produce.”

Why this purpose statement is effective:

• The subject of the proposal is clear: the establishment of a community garden.
• The specific goals of the project are outlined: promoting sustainable urban agriculture, fostering community engagement, and improving local access to fresh produce.
• The overall objective of the proposal is evident: securing funding and support.

Part 12 Purpose Statement Example For a Report

“The purpose of this report is to analyze current market trends in the electric vehicle (EV) industry, assess consumer preferences and buying behaviors, and provide strategic recommendations to guide [Company Name] in entering this growing market segment.”

• The subject of the report is provided: market trends in the electric vehicle industry.
• The specific goals of the report are analysis of market trends, assessment of consumer preferences, and strategic recommendations.
• The overall objective of the report is clear: providing guidance for the company’s entry into the EV market.

Part 13 Purpose Statement Example For a Project

“The purpose of this project is to design and implement a new employee wellness program that promotes physical and mental wellbeing in the workplace.”

This purpose statement clearly outlines the objective of the project, which is to create a new employee wellness program. The program is designed to promote physical and mental wellbeing in the workplace, which is a key concern for many employers. By implementing this program, the company aims to improve employee health, reduce absenteeism, and increase productivity. The purpose statement is concise and specific, providing a clear direction for the project team to follow. It highlights the importance of the project and its potential benefits for the company and its employees.

Part 14 Purpose Statement Templates

When writing a purpose statement, it can be helpful to use a template to ensure that you cover all the necessary components:

Template 1: To [action] [target audience] in order to [outcome]

This template is a straightforward way to outline your purpose statement. Simply fill in the blanks with the appropriate information:

• The purpose of […] is
• To [action]: What action do you want to take?
• [Target audience]: Who is your target audience?
• In order to [outcome]: What outcome do you hope to achieve?

For example:

• The purpose of our marketing campaign is to increase brand awareness among young adults in urban areas, in order to drive sales and revenue growth.
• The purpose of our employee training program is to improve customer service skills among our frontline staff, in order to enhance customer satisfaction and loyalty.
• The purpose of our new product launch is to expand our market share in the healthcare industry, by offering a unique solution to the needs of elderly patients with chronic conditions.

Template 2: This [project/product] is designed to [action] [target audience] by [method] in order to [outcome].

This template is useful for purpose statements that involve a specific project or product. Fill in the blanks with the appropriate information:

• This [project/product]: What is your project or product?
• Is designed to [action]: What action do you want to take?
• By [method]: What method will you use to achieve your goal?
• This app is designed to provide personalized nutrition advice to athletes by analyzing their training data in order to optimize performance.

Frequently Asked Questions

What are the key elements of a purpose statement.

A purpose statement should clearly communicate the main goal or objective of your writing. It should be concise and specific, providing a clear direction for your work. The key elements of a purpose statement include the topic or subject matter, the intended audience, and the overall goal or objective of your writing.

How can a purpose statement benefit your writing?

A purpose statement can help you stay focused and on track when writing. It can also help you to avoid going off-topic or getting bogged down in unnecessary details. By clearly identifying the main goal or objective of your writing, a purpose statement can help you to stay organized and ensure that your writing is effective and impactful.

What are some common mistakes to avoid when writing a purpose statement?

One common mistake is being too vague or general in your purpose statement. Another mistake is making your purpose statement too long or complex, which can make it difficult to understand. Additionally, it’s important to avoid including unnecessary information or details that are not directly relevant to your main goal or objective.

How can you tailor your purpose statement to your audience?

When writing a purpose statement, it’s important to consider your audience and their needs. You should tailor your purpose statement to your audience by using language and terminology that they will understand. You should also consider their level of knowledge or expertise on the subject matter and adjust your purpose statement accordingly.

What are some effective templates for writing a purpose statement?

There are many effective templates for writing a purpose statement, but one common approach is to use the following structure: “The purpose of this writing is to [insert goal or objective] for [insert audience] regarding [insert topic or subject matter].”

Can you provide examples of successful purpose statements?

• “The purpose of this report is to provide an analysis of the current market trends and make recommendations for future growth strategies for our company.”
• “The purpose of this essay is to explore the impact of social media on modern communication and its implications for society.”
• “The purpose of this proposal is to secure funding for a new community center that will provide educational and recreational opportunities for local residents.”
• 20 Inspiring Examples: How to Write a Personal Mission Statement
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How to Write a Qualitative Purpose Statement

Yashekia king.

A qualitative purpose statement in a research plan is an important step when you completing your master’s degree thesis or doctoral degree dissertation. The purpose statement allows you to clearly and concisely describe the intent of your qualitative study so that your reader can anticipate the information he will receive in your research report. Qualitative research involves capturing the complexity of behavior that occurs in natural settings, while the opposite goal of quantitative research is to collect data in the form of numbers. Write a qualitative purpose statement in about 100 words or less.

Describe the type of testing or method of inquiry you will use to do your qualitative research. State if you plan to perform an experiment or do a correlational analysis, the process of using statistical data to evaluate the extent of relations between variables. Explain if you will perform ethnographic research, a social science research method that depends on personal experience and participation rather than just observation.

Explain the purpose of the qualitative study you will perform. For instance, identify the variables you are testing and analyzing or the observable occurrences or phenomena you are striving to discover. Use words such as “discover,” “explore,” “develop” or “describe” regarding your research objective and indicate what is likely necessary to conduct the study. Foreshadow the hypotheses you will test or the questions your research will raise. Describe the significance of your study and how it will contribute to past research and existing knowledge on your research topic.

Name the setting and population of your qualitative study. For example, state that your research concerns employees working in a national restaurant chain or people with disabilities and their families. Describe how the researcher and participants will interact with one another in your study - if will they be observed, interviewed or asked to fill out a questionnaire, for example.

Expand on your primary qualitative study purpose by explaining additional purposes of your research. Describe extra variables related to these additional purposes as well as additional qualitative phenomena you want to discover. Reiterate the central concepts and ideas being studied.

• Read over your qualitative purpose statement, making sure the statement is written in future tense so that you indicate what the purpose and variables "will be." Check for errors in spelling, grammar, usage and punctuation.
• 1 WordNet: Correlational Analysis
• 2 American University: Notes on Qualitative Research; Julie Mertus
• 3 Emory University: The Elements of a Proposal; Frank Pajares; 2007

About the Author

YaShekia King, of Indianapolis, began writing professionally in 2003. Her work has appeared in several publications including the "South Bend Tribune" and "Clouds Across the Stars," an international book. She also is a licensed Realtor and clinical certified dental assistant. King holds a bachelor's degree in journalism from Ball State University.

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Definition of Data Interpretation

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21 Research Objectives Examples (Copy and Paste)

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Learn about our Editorial Process

Research objectives refer to the definitive statements made by researchers at the beginning of a research project detailing exactly what a research project aims to achieve.

These objectives are explicit goals clearly and concisely projected by the researcher to present a clear intention or course of action for his or her qualitative or quantitative study. 

Research objectives are typically nested under one overarching research aim. The objectives are the steps you’ll need to take in order to achieve the aim (see the examples below, for example, which demonstrate an aim followed by 3 objectives, which is what I recommend to my research students).

Research Objectives vs Research Aims

Research aim and research objectives are fundamental constituents of any study, fitting together like two pieces of the same puzzle.

The ‘research aim’ describes the overarching goal or purpose of the study (Kumar, 2019). This is usually a broad, high-level purpose statement, summing up the central question that the research intends to answer.

Example of an Overarching Research Aim:

“The aim of this study is to explore the impact of climate change on crop productivity.” 

Comparatively, ‘research objectives’ are concrete goals that underpin the research aim, providing stepwise actions to achieve the aim.

Objectives break the primary aim into manageable, focused pieces, and are usually characterized as being more specific, measurable, achievable, relevant, and time-bound (SMART).

Examples of Specific Research Objectives:

1. “To examine the effects of rising temperatures on the yield of rice crops during the upcoming growth season.” 2. “To assess changes in rainfall patterns in major agricultural regions over the first decade of the twenty-first century (2000-2010).” 3. “To analyze the impact of changing weather patterns on crop diseases within the same timeframe.”

The distinction between these two terms, though subtle, is significant for successfully conducting a study. The research aim provides the study with direction, while the research objectives set the path to achieving this aim, thereby ensuring the study’s efficiency and effectiveness.

How to Write Research Objectives

I usually recommend to my students that they use the SMART framework to create their research objectives.

SMART is an acronym standing for Specific, Measurable, Achievable, Relevant, and Time-bound. It provides a clear method of defining solid research objectives and helps students know where to start in writing their objectives (Locke & Latham, 2013).

Each element of this acronym adds a distinct dimension to the framework, aiding in the creation of comprehensive, well-delineated objectives.

Here is each step:

• Specific : We need to avoid ambiguity in our objectives. They need to be clear and precise (Doran, 1981). For instance, rather than stating the objective as “to study the effects of social media,” a more focused detail would be “to examine the effects of social media use (Facebook, Instagram, and Twitter) on the academic performance of college students.”
• Measurable: The measurable attribute provides a clear criterion to determine if the objective has been met (Locke & Latham, 2013). A quantifiable element, such as a percentage or a number, adds a measurable quality. For example, “to increase response rate to the annual customer survey by 10%,” makes it easier to ascertain achievement.
• Achievable: The achievable aspect encourages researchers to craft realistic objectives, resembling a self-check mechanism to ensure the objectives align with the scope and resources at disposal (Doran, 1981). For example, “to interview 25 participants selected randomly from a population of 100” is an attainable objective as long as the researcher has access to these participants.
• Relevance : Relevance, the fourth element, compels the researcher to tailor the objectives in alignment with overarching goals of the study (Locke & Latham, 2013). This is extremely important – each objective must help you meet your overall one-sentence ‘aim’ in your study.
• Time-Bound: Lastly, the time-bound element fosters a sense of urgency and prioritization, preventing procrastination and enhancing productivity (Doran, 1981). “To analyze the effect of laptop use in lectures on student engagement over the course of two semesters this year” expresses a clear deadline, thus serving as a motivator for timely completion.

You’re not expected to fit every single element of the SMART framework in one objective, but across your objectives, try to touch on each of the five components.

Research Objectives Examples

1. Field: Psychology

Aim: To explore the impact of sleep deprivation on cognitive performance in college students.

• Objective 1: To compare cognitive test scores of students with less than six hours of sleep and those with 8 or more hours of sleep.
• Objective 2: To investigate the relationship between class grades and reported sleep duration.
• Objective 3: To survey student perceptions and experiences on how sleep deprivation affects their cognitive capabilities.

2. Field: Environmental Science

Aim: To understand the effects of urban green spaces on human well-being in a metropolitan city.

• Objective 1: To assess the physical and mental health benefits of regular exposure to urban green spaces.
• Objective 2: To evaluate the social impacts of urban green spaces on community interactions.
• Objective 3: To examine patterns of use for different types of urban green spaces. 

3. Field: Technology

Aim: To investigate the influence of using social media on productivity in the workplace.

• Objective 1: To measure the amount of time spent on social media during work hours.
• Objective 2: To evaluate the perceived impact of social media use on task completion and work efficiency.
• Objective 3: To explore whether company policies on social media usage correlate with different patterns of productivity.

4. Field: Education

Aim: To examine the effectiveness of online vs traditional face-to-face learning on student engagement and achievement.

• Objective 1: To compare student grades between the groups exposed to online and traditional face-to-face learning.
• Objective 2: To assess student engagement levels in both learning environments.
• Objective 3: To collate student perceptions and preferences regarding both learning methods.

5. Field: Health

Aim: To determine the impact of a Mediterranean diet on cardiac health among adults over 50.

• Objective 1: To assess changes in cardiovascular health metrics after following a Mediterranean diet for six months.
• Objective 2: To compare these health metrics with a similar group who follow their regular diet.
• Objective 3: To document participants’ experiences and adherence to the Mediterranean diet.

6. Field: Environmental Science

Aim: To analyze the impact of urban farming on community sustainability.

• Objective 1: To document the types and quantity of food produced through urban farming initiatives.
• Objective 2: To assess the effect of urban farming on local communities’ access to fresh produce.
• Objective 3: To examine the social dynamics and cooperative relationships in the creating and maintaining of urban farms.

7. Field: Sociology

Aim: To investigate the influence of home offices on work-life balance during remote work.

• Objective 1: To survey remote workers on their perceptions of work-life balance since setting up home offices.
• Objective 2: To conduct an observational study of daily work routines and family interactions in a home office setting.
• Objective 3: To assess the correlation, if any, between physical boundaries of workspaces and mental boundaries for work in the home setting.

8. Field: Economics

Aim: To evaluate the effects of minimum wage increases on small businesses.

• Objective 1: To analyze cost structures, pricing changes, and profitability of small businesses before and after minimum wage increases.
• Objective 2: To survey small business owners on the strategies they employ to navigate minimum wage increases.
• Objective 3: To examine employment trends in small businesses in response to wage increase legislation.

9. Field: Education

Aim: To explore the role of extracurricular activities in promoting soft skills among high school students.

• Objective 1: To assess the variety of soft skills developed through different types of extracurricular activities.
• Objective 2: To compare self-reported soft skills between students who participate in extracurricular activities and those who do not.
• Objective 3: To investigate the teachers’ perspectives on the contribution of extracurricular activities to students’ skill development.

10. Field: Technology

Aim: To assess the impact of virtual reality (VR) technology on the tourism industry.

• Objective 1: To document the types and popularity of VR experiences available in the tourism market.
• Objective 2: To survey tourists on their interest levels and satisfaction rates with VR tourism experiences.
• Objective 3: To determine whether VR tourism experiences correlate with increased interest in real-life travel to the simulated destinations.

11. Field: Biochemistry

Aim: To examine the role of antioxidants in preventing cellular damage.

• Objective 1: To identify the types and quantities of antioxidants in common fruits and vegetables.
• Objective 2: To determine the effects of various antioxidants on free radical neutralization in controlled lab tests.
• Objective 3: To investigate potential beneficial impacts of antioxidant-rich diets on long-term cellular health.

12. Field: Linguistics

Aim: To determine the influence of early exposure to multiple languages on cognitive development in children.

• Objective 1: To assess cognitive development milestones in monolingual and multilingual children.
• Objective 2: To document the number and intensity of language exposures for each group in the study.
• Objective 3: To investigate the specific cognitive advantages, if any, enjoyed by multilingual children.

13. Field: Art History

Aim: To explore the impact of the Renaissance period on modern-day art trends.

• Objective 1: To identify key characteristics and styles of Renaissance art.
• Objective 2: To analyze modern art pieces for the influence of the Renaissance style.
• Objective 3: To survey modern-day artists for their inspirations and the influence of historical art movements on their work.

14. Field: Cybersecurity

Aim: To assess the effectiveness of two-factor authentication (2FA) in preventing unauthorized system access.

• Objective 1: To measure the frequency of unauthorized access attempts before and after the introduction of 2FA.
• Objective 2: To survey users about their experiences and challenges with 2FA implementation.
• Objective 3: To evaluate the efficacy of different types of 2FA (SMS-based, authenticator apps, biometrics, etc.).

15. Field: Cultural Studies

Aim: To analyze the role of music in cultural identity formation among ethnic minorities.

• Objective 1: To document the types and frequency of traditional music practices within selected ethnic minority communities.
• Objective 2: To survey community members on the role of music in their personal and communal identity.
• Objective 3: To explore the resilience and transmission of traditional music practices in contemporary society.

16. Field: Astronomy

Aim: To explore the impact of solar activity on satellite communication.

• Objective 1: To categorize different types of solar activities and their frequencies of occurrence.
• Objective 2: To ascertain how variations in solar activity may influence satellite communication.
• Objective 3: To investigate preventative and damage-control measures currently in place during periods of high solar activity.

17. Field: Literature

Aim: To examine narrative techniques in contemporary graphic novels.

• Objective 1: To identify a range of narrative techniques employed in this genre.
• Objective 2: To analyze the ways in which these narrative techniques engage readers and affect story interpretation.
• Objective 3: To compare narrative techniques in graphic novels to those found in traditional printed novels.

18. Field: Renewable Energy

Aim: To investigate the feasibility of solar energy as a primary renewable resource within urban areas.

• Objective 1: To quantify the average sunlight hours across urban areas in different climatic zones. 
• Objective 2: To calculate the potential solar energy that could be harnessed within these areas.
• Objective 3: To identify barriers or challenges to widespread solar energy implementation in urban settings and potential solutions.

19. Field: Sports Science

Aim: To evaluate the role of pre-game rituals in athlete performance.

• Objective 1: To identify the variety and frequency of pre-game rituals among professional athletes in several sports.
• Objective 2: To measure the impact of pre-game rituals on individual athletes’ performance metrics.
• Objective 3: To examine the psychological mechanisms that might explain the effects (if any) of pre-game ritual on performance.

20. Field: Ecology

Aim: To investigate the effects of urban noise pollution on bird populations.

• Objective 1: To record and quantify urban noise levels in various bird habitats.
• Objective 2: To measure bird population densities in relation to noise levels.
• Objective 3: To determine any changes in bird behavior or vocalization linked to noise levels.

21. Field: Food Science

Aim: To examine the influence of cooking methods on the nutritional value of vegetables.

• Objective 1: To identify the nutrient content of various vegetables both raw and after different cooking processes.
• Objective 2: To compare the effect of various cooking methods on the nutrient retention of these vegetables.
• Objective 3: To propose cooking strategies that optimize nutrient retention.

The Importance of Research Objectives

The importance of research objectives cannot be overstated. In essence, these guideposts articulate what the researcher aims to discover, understand, or examine (Kothari, 2014).

When drafting research objectives, it’s essential to make them simple and comprehensible, specific to the point of being quantifiable where possible, achievable in a practical sense, relevant to the chosen research question, and time-constrained to ensure efficient progress (Kumar, 2019). 

Remember that a good research objective is integral to the success of your project, offering a clear path forward for setting out a research design , and serving as the bedrock of your study plan. Each objective must distinctly address a different dimension of your research question or problem (Kothari, 2014). Always bear in mind that the ultimate purpose of your research objectives is to succinctly encapsulate your aims in the clearest way possible, facilitating a coherent, comprehensive and rational approach to your planned study, and furnishing a scientific roadmap for your journey into the depths of knowledge and research (Kumar, 2019). 

Kothari, C.R (2014). Research Methodology: Methods and Techniques . New Delhi: New Age International.

Kumar, R. (2019). Research Methodology: A Step-by-Step Guide for Beginners .New York: SAGE Publications.

Doran, G. T. (1981). There’s a S.M.A.R.T. way to write management’s goals and objectives. Management review, 70 (11), 35-36.

Locke, E. A., & Latham, G. P. (2013). New Developments in Goal Setting and Task Performance . New York: Routledge.

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Free Research Purpose Statement Generator for Students

What is a good research purpose statement? Below, you’ll find its definition, writing tips, and some excellent examples. Keep reading!

• ️✅ How to Use the Tool
• ️🔬 Research Purpose Statement Basics
• ️🖊️ How to Write It
• ️💪 Tips on Writing a Strong Statement
• ️🤩 Free Examples
• ️🔗 References

✅ Research Purpose Statement Generator: How to Use

A research purpose statement is an essential part of any research. It helps you stay focused on the problem you’re investigating and also catches your reader’s attention.

However, it might take some time and energy to develop a solid purpose statement. Our goal is to make your studies more enjoyable with the help of our tool!

Here’s how our research purpose statement generator works:

• Choose the type of your research.
• Choose a verb that best describes your study’s purpose.
• Type in your research group, the object of your study , and what is impacted by it.
• Provide the information on the place and time of the research.
• Describe the research group 2 if necessary.

🔬 Research Purpose Statement: the Basics

Research purpose definition.

A research purpose statement describes your research’s reason, goal, and process. It explains what the research explores, how it does it, and where it takes place. A research purpose is simple and concise. It is usually placed at the end of the introduction.

The Importance of Research Purpose Statement

A research purpose statement helps your reader understand what the research will be about and explains the significance of your study . It also helps you avoid distractions and focus on the relevant information needed for the research.

Research Purpose Statement vs. Thesis Statement

The purpose of both types of statements is to let readers know what to expect from your work. But here’s a difference:

• A thesis statement "> A thesis statement makes a claim and predicts how the essay will unfold.
• A purpose statement explains what the research will be about, how it will be conducted, and where it will take place.

🖊️ How to Write a Research Purpose Statement

Need to write your research purpose statement but don’t know where to start? We’ve got your back! Here, we explain step by step how to make a solid purpose statement for your research.

Step 1 – Identify Whether Your Study Is Quantitative or Qualitative

Research purpose statements vary depending on the type of research. One in a quantitative paper should focus on specific numbers. A qualitative research purpose statement focuses on precise intent. You can start your statement with, “The purpose of this qualitative/quantitative research is…”

Step 2 – Define the Research Process

Explain the methods you are going to use in your research. What data or instruments are you going to use? By explaining your research process , you will make it easier for the reader to understand what to expect from your work.

Step 3 – Explain What Will Be Researched and Where It Will Take Place

Try to provide enough background information for your reader. Where is the research going to take place? Does the reader need to know about the population of that location?

Step 4 – Make a Statement

Now, put everything together and create your statement.

💪 Tips on Making a Great Research Purpose Statement

• Write your research purpose statement together with the problem statement . This will ensure that the purpose is connected to the problem.
• Write only one purpose statement. Your purpose should cover the entire research, so there’s no need to divide it into smaller parts.
• Avoid including too many details. The purpose statement should be easy to understand. Otherwise, your reader might get lost.
• Use simple words. Don’t go overboard with creativity because it might distract you from the problem.

🤩 Research Purpose Statement Examples

Want some inspiration for your research purpose statement? Here are some of the best examples to help you out!

Quantitative research:

This quantitative research aims to compare the academic results of the students attending online classes and those attending traditional classes regarding exam scores and GPA.

The purpose above is short and clear. It lets the reader know what the research is going to be about.

Qualitative research:

The purpose of this qualitative research is to understand how mental health education impacts the academic achievements of high school students.

This purpose is also short and precise. It doesn’t have too many details but is straight to the point.

We hope this article was helpful. Remember that you can always use our research purpose statement generator for free. Make sure to check our APA title page generator and research question maker too!

❓ Research Purpose Statement FAQ

❓ what is purpose statement in research.

A purpose statement explains to your reader what your research will be about. It should describe the goal, methods, and place of your study. It is usually at the end of the introduction part.

❓ How to write a purpose statement for a research paper?

To write a purpose statement for a research paper, you should follow these steps:

• Identify the type of your research
• Explain the process of the research
• Describe who or what you are going to research and where it takes place
• Finally, put it all together

❓ What is a purpose statement in research example?

“The purpose of this quantitative research is to compare the academic results of the students attending online classes and those attending traditional classes regarding exam scores and GPA.” This quantitative research purpose statement is short and clear. It only provides a little information but lets the reader know what to expect from the research.

❓ Where does the purpose statement go in a research paper?

A purpose statement is usually at the end of the introductory part of your research. Since its function is to tell your reader what to expect from your paper, you should place it before your study’s main body.

Updated: Aug 23rd, 2024

🔗 References

• Research Paper Purpose Statement Examples: Your Dictionary
• Thesis and Purpose Statements: University of Wisconsin
• Writing Effective Purpose Statements: University of Washington
• Importance of a Purpose Statement in Research: The Classroom

Stating the Obvious: Writing Assumptions, Limitations, and Delimitations

During the process of writing your thesis or dissertation, you might suddenly realize that your research has inherent flaws. Don’t worry! Virtually all projects contain restrictions to your research. However, being able to recognize and accurately describe these problems is the difference between a true researcher and a grade-school kid with a science-fair project. Concerns with truthful responding, access to participants, and survey instruments are just a few of examples of restrictions on your research. In the following sections, the differences among delimitations, limitations, and assumptions of a dissertation will be clarified.

Delimitations

Delimitations are the definitions you set as the boundaries of your own thesis or dissertation, so delimitations are in your control. Delimitations are set so that your goals do not become impossibly large to complete. Examples of delimitations include objectives, research questions, variables, theoretical objectives that you have adopted, and populations chosen as targets to study. When you are stating your delimitations, clearly inform readers why you chose this course of study. The answer might simply be that you were curious about the topic and/or wanted to improve standards of a professional field by revealing certain findings. In any case, you should clearly list the other options available and the reasons why you did not choose these options immediately after you list your delimitations. You might have avoided these options for reasons of practicality, interest, or relativity to the study at hand. For example, you might have only studied Hispanic mothers because they have the highest rate of obese babies. Delimitations are often strongly related to your theory and research questions. If you were researching whether there are different parenting styles between unmarried Asian, Caucasian, African American, and Hispanic women, then a delimitation of your study would be the inclusion of only participants with those demographics and the exclusion of participants from other demographics such as men, married women, and all other ethnicities of single women (inclusion and exclusion criteria). A further delimitation might be that you only included closed-ended Likert scale responses in the survey, rather than including additional open-ended responses, which might make some people more willing to take and complete your survey. Remember that delimitations are not good or bad. They are simply a detailed description of the scope of interest for your study as it relates to the research design. Don’t forget to describe the philosophical framework you used throughout your study, which also delimits your study.

Limitations

Limitations of a dissertation are potential weaknesses in your study that are mostly out of your control, given limited funding, choice of research design, statistical model constraints, or other factors. In addition, a limitation is a restriction on your study that cannot be reasonably dismissed and can affect your design and results. Do not worry about limitations because limitations affect virtually all research projects, as well as most things in life. Even when you are going to your favorite restaurant, you are limited by the menu choices. If you went to a restaurant that had a menu that you were craving, you might not receive the service, price, or location that makes you enjoy your favorite restaurant. If you studied participants’ responses to a survey, you might be limited in your abilities to gain the exact type or geographic scope of participants you wanted. The people whom you managed to get to take your survey may not truly be a random sample, which is also a limitation. If you used a common test for data findings, your results are limited by the reliability of the test. If your study was limited to a certain amount of time, your results are affected by the operations of society during that time period (e.g., economy, social trends). It is important for you to remember that limitations of a dissertation are often not something that can be solved by the researcher. Also, remember that whatever limits you also limits other researchers, whether they are the largest medical research companies or consumer habits corporations. Certain kinds of limitations are often associated with the analytical approach you take in your research, too. For example, some qualitative methods like heuristics or phenomenology do not lend themselves well to replicability. Also, most of the commonly used quantitative statistical models can only determine correlation, but not causation.

Assumptions

Assumptions are things that are accepted as true, or at least plausible, by researchers and peers who will read your dissertation or thesis. In other words, any scholar reading your paper will assume that certain aspects of your study is true given your population, statistical test, research design, or other delimitations. For example, if you tell your friend that your favorite restaurant is an Italian place, your friend will assume that you don’t go there for the sushi. It’s assumed that you go there to eat Italian food. Because most assumptions are not discussed in-text, assumptions that are discussed in-text are discussed in the context of the limitations of your study, which is typically in the discussion section. This is important, because both assumptions and limitations affect the inferences you can draw from your study. One of the more common assumptions made in survey research is the assumption of honesty and truthful responses. However, for certain sensitive questions this assumption may be more difficult to accept, in which case it would be described as a limitation of the study. For example, asking people to report their criminal behavior in a survey may not be as reliable as asking people to report their eating habits. It is important to remember that your limitations and assumptions should not contradict one another. For instance, if you state that generalizability is a limitation of your study given that your sample was limited to one city in the United States, then you should not claim generalizability to the United States population as an assumption of your study. Statistical models in quantitative research designs are accompanied with assumptions as well, some more strict than others. These assumptions generally refer to the characteristics of the data, such as distributions, correlational trends, and variable type, just to name a few. Violating these assumptions can lead to drastically invalid results, though this often depends on sample size and other considerations.

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3+ SAMPLE Qualitative Research Statement in PDF

Qualitative research statement, 3+ sample qualitative research statement, what is a qualitative research statement, types of qualitative research statement, steps in writing a qualitative research statement, how to write a qualitative research proposal, what are the 5 qualitative research types, how do quantitative and qualitative methods differ, what are mixed methods research, what’s the difference between validity and reliability, what is the purpose of hypothesis testing.

Qualitative Descriptive Study Research Statement

Qualitative Research Purpose Statement

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Sample Qualitative Research Statement

1. include the problem to be solved, 2. framework for theoretical considerations, 3. methods of investigation, 4. appendices and references are included, share this post on your network, you may also like these articles, bank statement.

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FREE 3+ Qualitative Research Statement Samples [ Purpose, Problem, Positionality ]

As a humanistic or idealistic method, qualitative research focuses on understanding a study query. People’s views, experiences, attitudes, behavior , and interactions are studied using qualitative methods. People and organizations are thus researched in their natural environment. If you’re currently starting a research study , you would know that one of the processes in a qualitative paper is writing the research statement. For your convenience, we offer you free and ready-to-use samples of a Qualitative Research Statement that you could use to present your research’s purpose or problem. Keep on reading to find out more.

Qualitative Research Statement

3+ qualitative research statement samples, 1. qualitative descriptive study research statement, 2. qualitative research paper problem statement, 3. qualitative research purpose statement, 4. qualitative project research statement, what is a qualitative research statement, how to make a qualitative research statement, 1. identify the type of testing or inquiry approach you’ll employ to conduct your qualitative research., 2. describe the goal of the qualitative research you’ll do., 3. give your qualitative study a name and a target population., 4. extend the initial purpose of your qualitative study by describing your research’s additional objectives., what’s the difference between a qualitative research statement and a quantitative research statement, what is the definition of a research purpose statement, in qualitative research, how are sources used, why would you select qualitative research over quantitative research.

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A distinct objective or intent will be presented in qualitative research purpose statements, and a specific idea will be investigated. You’ll also be able to see exactly how and where you’ll be learning. A solid qualitative purpose statement includes details on the study’s central phenomenon and participants. Rather of focusing on countable, numerical figures, the data is descriptive. A qualitative research statement explains what is being investigated or examined, how it is being investigated, and where it is being investigated. Essentially, a qualitative research purpose statements, which investigate a central phenomenon.

If you’re interested in making a qualitative research statement, you could use the free templates provided above to help you out. Other than that, here are the following statements you could use to write your statement from the ground up:

Indicate if you intend to conduct an experiment or conduct a correlational study, which is the act of evaluating the extent of relationships between variables using statistical data. Explain whether you plan to conduct ethnographic research, which is a type of social science research that relies on personal experience and engagement rather than just observation.

Determine the variables you’ll be testing and evaluating, as well as the observable occurrences or phenomena you’ll be looking for. Predict which hypotheses you’ll test and which questions your study will raise. Describe the significance of your research and how it will add to previous research and understanding on your issue.

Describe how the researcher and participants will interact in your study, such as if they will be observed, interviewed, or asked to complete a questionnaire. Declare, for example, that your study is focused on a specific population or demography.

Describe any other variables that are relevant to these additional aims, as well as any qualitative phenomena you want to investigate. Reiterate the key concepts and ideas that you’ve learned thus far.

Quantitative research is concerned with numbers and figures, whereas qualitative research is concerned with words and their interpretations. Quantitative approaches allow you to test a theory by gathering and analyzing data in a methodical way, whereas qualitative methods allow you to dig deeper into ideas and experiences.

A purpose statement is a declarative statement that highlights the major goal or goals of a research study. A purpose statement acts as an introduction to the resulting article or dissertation chapter and provides some direction in selecting a research question.

Qualitative researchers collect data from a variety of sources in order to gain a better understanding of the subject they are researching. In-depth interviews, focus groups, standardized interviews, and artifacts such as books or works of art are among the data sources.

Qualitative research methods provide a dynamic approach to study, allowing the researcher to follow up on responses made by respondents in real-time, resulting in important dialogue around a topic.

Overall, a research statement is used to determine the substance of a research paper, the research’s primary themes, and the research’s future direction. To assist you with this, you may download our easily modifiable and printable samples of Qualitative Research Statements today!

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Problem and Purpose Statements for a Qualitative Study

Find three articles (3) related to the topic of improving student learning outcomes by teaching brain-based learning to educators through professional development training.

Develop a 1- to 2-paragraph PROBLEM statement that is the result of a review of the three (3) articles you located on your topic. In the paragraphs clearly answer the following: Briefly describe the phenomena you are interested in studying. Briefly summarize the key findings or what is understood about this phenomena based on the three articles you reviewed. Briefly identify the “gap”—what do you see as an important, relevant, next step in learning more about this topic that would be appropriate for a qualitative study. 3. Write a PURPOSE statement using terminology of qualitative research. The purpose statement should contain: A statement using the following template: The purpose of this qualitative study is to [choose one: explore, describe, understand, explain] the meaning/experience/culture/stories of [phenomenon of interest] in [population/setting]. A justification of why the purpose of your study is suited for the qualitative approach in terms of: The phenomena you choose -The “fit” with a constructivist epistemology and ontology -The relevance of the naturalistic setting or context to the phenomena of interest Based on your knowledge of reflexivity, explain your relationship to the problem; and the issues of bias and positionality to be addressed. Include a brief description of the setting (P-20 classrooms) and possible sources of data. 4. Write a RESEARCH QUESTION using terminology of qualitative research. The research question should incorporate and mirror the purpose statement using the following template: What is the meaning/experience/culture/stories of [phenomenon of interest] in [population/setting]? 5. Use APA 7 in articles from the last five (5) years 6. Write annotated bibliography from articles used.

Topic References:

improving learning outcomes for students by teaching brain based learning to teachers – Bing

– improving learning outcomes for students by teaching brain based learning in Professional Development Trainings for educators – Bing

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Example of Purpose of the Study: Writing Your Purpose Statement

• Author Dr. Robertson Prime, Research Fellow
• Published April 1, 2024

Table of Contents

Example of Purpose of the Study or Purpose Statement

What is a purpose of the study in a single sentence.

A purpose of the study statement succinctly explains the specific intent or objective that the research aims to achieve in a single, focused sentence. This must be present in all types of research whether, quantitative or qualitative research. The statement explains why the study will be conducted, or the primary reason for conducting the study.

Purpose of the Study

A purpose statement must be well-defined in your dissertation. Please learn more from our blog on dissertation template . This blog will provide you with an example of purpose of the study.

Do not confuse this section with the problem statement; they are two different sections of an introduction chapter of a dissertation

Example of Purpose Statement Overview

Learn more from this example of purpose of the study.

Here is an example of purpose statement overview:

“The purpose of this qualitative phenomenological study is to explore the lived experiences of first-generation college students at a large public university in overcoming barriers to academic success.”

In this statement:

• It clearly states the intent is “to explore” using qualitative, phenomenological methods.
• It specifies the key phenomenon of interest as the “lived experiences” of the participants.
• It identifies the specific population as “first-generation college students.”
• It bounds the scope to a “large public university” setting.
• The purpose connects to the issue of “overcoming barriers to academic success.”

This succinct statement covers the what (explore lived experiences), who (first-gen students), where (large public university), and why (overcoming barriers) in one focused sentence. The language is clear, direct, and aligns well with the qualitative phenomenological approach proposed.

This is an example of purpose of the study formulated by our team of writers at bestdissertationwriter.com . Ask us to help you with your doctoral dissertation or any other form of dissertation writing services, and we will do it affordable rate, JUST FOR YOU.

Mistakes People Make When Writing Their Purpose Statement

Here are some common mistakes people make when writing their purpose statement for a research paper:

• Making the purpose statement too long instead of concise.
• Stating research methods or approaches instead of the purpose.
• Describing what was studied rather than the intent/objective or research question .
• Including multiple purposes in one statement.
• Failing to connect the purpose to the research problem/questions.
• Writing the purpose as a question rather than a statement.
• Stating a purpose disconnected from the literature/theory.
• Failing to align the purpose with the chosen methodology.

What are the Best Practices for Writing your Purpose Statement?

Here are the 5 key best practices for writing an effective purpose statement:

• Use clear, concise language to state the primary objective /intent in one focused sentence and should be consistent with the research question.
• Directly align the purpose with your research questions/hypotheses and identified gap/problem.
• Highlight the core concepts, variables, or relationships to be examined, without excessive detail.
• Ensure the stated purpose is specific yet realistic given your research scope and methods.
• Frame the purpose in a way that allows for potential unanticipated findings or implications.

Example of Purpose of the Study

Example of Purpose of the Study Template for Your Research

Here is a template you can use for crafting a clear and effective purpose statement for a research paper:

“The purpose of this [TYPE OF STUDY] is to [EXAMINE/EXPLORE/DESCRIBE/ETC.] the [KEY CONCEPT/PHENOMENON/RELATIONSHIP] among/between [PARTICIPANTS/SUBJECTS/VARIABLES] in [CONTEXT/SETTING] in order to [IDENTIFY/UNDERSTAND/DETERMINE/ETC.] [RESEARCH PROBLEM/ISSUE/GAP].”

Some examples following this template:

• “The purpose of this quantitative correlational study is to examine the relationship between social media usage and academic performance among high school students in order to understand factors impacting student achievement.”
• “The purpose of this mixed methods case study is to explore the implementation and effectiveness of a new reading intervention program for elementary students in an urban school district in order to identify best practices.”
• “The purpose of this experimental research is to determine the effects of mindfulness training on stress reduction and well-being in healthcare workers in order to develop organizational support strategies.”

The key components are:

• Academic verb – examine/explore/describe/etc.
• Core concepts/variables
• Participant/subject details
• Context/setting
• Stated intent – identify/understand/determine/etc.
• Research problem/issue/gap

Feel free to adapt this template to suit your specific research aims or research question while still capturing the essential purpose elements in one coherent statement.

Example of Purpose of the Study or Sample Purpose Statement

Here is an example of purpose of the study or sample purpose statement. With this example of purpose of the study, you can equip yourself with appropriate skills required for developing such statements for your research.

The purpose of this explanatory mixed methods study is to investigate the factors contributing to employee burnout and low job satisfaction among nurses in emergency departments (EDs) at urban hospitals. Specifically, this study aims to explore the lived experiences and perceptions of ED nurses regarding workplace stressors, coping strategies, and organizational support structures through qualitative interviews and open-ended surveys. Additionally, it will examine quantitative relationships between key variables such as nurse-patient ratios, overtime hours, years of experience, and standardized measures of burnout, job satisfaction, and turnover intentions. By combining insights from both qualitative and quantitative data, this research endeavors to develop a comprehensive understanding of the underlying causes and potential interventions to address ED nurse burnout. The findings may inform evidence-based policies and practices to improve nurse well-being, retention, and quality of patient care in high-stress healthcare environments.

This statement clearly specifies the methodology as mixed methods, purpose of your study, the core concepts involved, the study population of ED nurses, the research context of urban hospitals, and the overall intent to investigate factors around burnout using multiple approaches. It connects the purpose to identifying interventions for an important issue in nursing and healthcare. This can act as an example of quantitative research purpose statement.

FAQ – Example of Purpose of the Study or Purpose Statement

What is a purpose statement.

A purpose statement is a clear and concise description of the goals and objectives of a research study. It outlines the specific reasons why the study is being conducted. This can be qualitative or quantitative purpose statements as it explains the primary purpose of your research. Learn more from this example of purpose of the study.

How is the purpose of the study different from a research question?

The purpose of the study is a broader explanation of why the research is being undertaken, while a research question is a specific query that the study seeks to answer. It is important that you know the difference between problem statement and purpose statement.

What is the importance of writing the purpose statement?

Writing your purpose statement is crucial as it provides direction and clarity to your research. It helps in outlining the scope and focus of the study.

Can you provide an example of a qualitative purpose statement?

An example of a qualitative purpose statement could be: “The purpose of this qualitative study is to explore the lived experiences of cancer survivors.”

How should a purpose statement be structured in a quantitative research study?

The purpose statement in a quantitative research study should clearly define the goals, objectives, and rationale of the study, focusing on measurable outcomes. The purpose statement should focus on the primary goal of the study, generally a focus of your research.

What role does the purpose section play in a research project?

The purpose section of a research project is where you articulate the overall aim and significance of the study, setting the context for the research that will follow.

What should researchers keep in mind as to the purpose of their research?

Researchers should keep in mind the purpose of their research throughout the study to stay focused on their objectives and ensure that the research remains aligned with the intended goals.

How can one ensure that their purpose statement is well-defined?

To ensure that your research has a well-defined purpose statement , it is important to clearly outline the research aims, objectives, and expected outcomes at the outset of the study.

What is the relationship between a problem statement and a purpose statement?

A problem statement identifies the research issue or gap that needs to be addressed, while the purpose statement explicitly states the intent or objective of the study in attempting to address that particular problem or gap.

Dr. Robertson Prime, Research Fellow

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• Open access
• Published: 28 August 2024

A qualitative study identifying implementation strategies using the i-PARIHS framework to increase access to pre-exposure prophylaxis at federally qualified health centers in Mississippi

• Trisha Arnold   ORCID: orcid.org/0000-0003-3556-5717 1 , 2 ,
• Laura Whiteley 2 ,
• Kayla K. Giorlando 1 ,
• Andrew P. Barnett 1 , 2 ,
• Ariana M. Albanese 2 ,
• Avery Leigland 1 ,
• Courtney Sims-Gomillia 3 ,
• A. Rani Elwy 2 , 5 ,
• Precious Patrick Edet 3 ,
• Demetra M. Lewis 4 ,
• James B. Brock 4 &
• Larry K. Brown 1 , 2  

Implementation Science Communications volume  5 , Article number:  92 ( 2024 ) Cite this article

3 Altmetric

Metrics details

Mississippi (MS) experiences disproportionally high rates of new HIV infections and limited availability of pre-exposure prophylaxis (PrEP). Federally Qualified Health Centers (FQHCs) are poised to increase access to PrEP. However, little is known about the implementation strategies needed to successfully integrate PrEP services into FQHCs in MS.

The study had two objectives: identify barriers and facilitators to PrEP use and to develop tailored implementation strategies for FQHCs.

Semi-structured interviews were conducted with 19 staff and 17 PrEP-eligible patients in MS FQHCs between April 2021 and March 2022. The interview was guided by the integrated-Promoting Action on Research Implementation in Health Services (i-PARIHS) framework which covered PrEP facilitators and barriers. Interviews were coded according to the i-PARIHS domains of context, innovation, and recipients, followed by thematic analysis of these codes. Identified implementation strategies were presented to 9 FQHC staff for feedback.

Data suggested that PrEP use at FQHCs is influenced by patient and clinic staff knowledge with higher levels of knowledge reflecting more PrEP use. Perceived side effects are the most significant barrier to PrEP use for patients, but participants also identified several other barriers including low HIV risk perception and untrained providers. Despite these barriers, patients also expressed a strong motivation to protect themselves, their partners, and their communities from HIV. Implementation strategies included education and provider training which were perceived as acceptable and appropriate.

Conclusions

Though patients are motivated to increase protection against HIV, multiple barriers threaten uptake of PrEP within FQHCs in MS. Educating patients and providers, as well as training providers, are promising implementation strategies to overcome these barriers.

Peer Review reports

Contributions to the literature

We propose utilizing Federally Qualified Health Centers (FQHCs) to increase pre-exposure prophylaxis (PrEP) use among people living in Mississippi.

Little is currently known about how to distribute PrEP at FQHCs.

We comprehensively describe the barriers and facilitators to implementing PrEP at FQHCs.

Utilizing effective implementation strategies of PrEP, such as education and provider training at FQHCs, may increase PrEP use and decrease new HIV infections.

Introduction

The HIV outbreak in Mississippi (MS) is among the most critical in the United States (U.S.). It is distinguished by significant inequalities, a considerable prevalence of HIV in remote areas, and low levels of HIV medical care participation and virologic suppression [ 1 ]. MS has consistently ranked among the states with the highest HIV rates in the U.S. This includes being the 6th highest in new HIV diagnoses [ 2 ] and 2nd highest in HIV diagnoses among men who have sex with men (MSM) compared to other states [ 2 , 3 , 4 ]. Throughout MS, the HIV epidemic disproportionately affects racial and ethnic minority groups, particularly among Black individuals. A spatial epidemiology and statistical modeling study completed in MS identified HIV hot spots in the MS Delta region, Southern MS, and in greater Jackson, including surrounding rural counties [ 5 ]. Black race and urban location were positively associated with HIV clusters. This disparity is often driven by the complex interplay of social, economic, and structural factors, including poverty, limited access to healthcare, and stigma [ 5 ].

Pre-exposure prophylaxis (PrEP) has gained significant recognition due to its safety and effectiveness in preventing HIV transmission when taken as prescribed [ 6 , 7 , 8 , 9 ]. However, despite the progression in PrEP and its accessibility, its uptake has been slow among individuals at high risk of contracting HIV, particularly in Southern states such as MS [ 10 , 11 , 12 , 13 , 14 ]. According to the CDC [ 5 ], “4,530 Mississippians at high risk for HIV could potentially benefit from PrEP, but only 927 were prescribed PrEP.” Several barriers hinder PrEP use in MS including limited access to healthcare, cost, stigma, and medical mistrust [ 15 , 16 , 17 ].

Federally qualified health centers (FQHCs) are primary healthcare organizations that are community-based and patient-directed, serve geographically and demographically diverse patients with limited access to medical care, and provide care regardless of a patient’s ability to pay [ 18 ]. FQHCs in these areas exhibit reluctance in prescribing or counseling patients regarding PrEP, primarily because they lack the required training and expertise [ 19 , 20 , 21 ]. Physicians in academic medical centers are more likely to prescribe PrEP compared to those in community settings [ 22 ]. Furthermore, providers at FQHCs may exhibit less familiarity with conducting HIV risk assessments, express concerns regarding potential side effects of PrEP, and have mixed feelings about prescribing it [ 23 , 24 ]. Task shifting might also be needed as some FQHCs may lack sufficient physician support to manage all aspects of PrEP care. Tailored strategies and approaches are necessary for FQHCs to effectively navigate the many challenges that threaten their patients’ access to and utilization of PrEP.

The main objectives of this study were to identify the barriers and facilitators to PrEP use and to develop tailored implementation strategies for FQHCs providing PrEP. To service these objectives, this study had three specific aims. Aim 1 involved conducting a qualitative formative evaluation guided by the integrated-Promoting Action on Research Implementation in Health Services (i-PARIHS) framework- with FQHC staff and PrEP-eligible patients across three FQHCs in MS [ 25 ]. Interviews covered each of the three i-PARIHS domains: context, innovation, and recipients. These interviews sought to identify barriers and facilitators to implementing PrEP. Aim 2 involved using interview data to select and tailor implementation strategies from the Expert Recommendations for Implementing Change (ERIC) project [ 26 ] (e.g., provider training) and methods (e.g., telemedicine, PrEP navigators) for the FQHCs. Aim 3 was to member-check the selected implementation strategies and further refine these if necessary. Data from all three aims are presented below. The standards for reporting qualitative research (SRQR) checklist was used to improve the transparency of reporting this qualitative study [ 27 ].

Formative evaluation interviews

Interviews were conducted with 19 staff and 17 PrEP-eligible patients from three FQHCs in Jackson, Canton, and Clarksdale, Mississippi. Staff were eligible to participate if they were English-speaking and employed by their organization for at least a year. Eligibility criteria for patients included: 1) English speaking, 2) aged 18 years or older, 3) a present or prior patient at the FQHC, 4) HIV negative, and 5) currently taking PrEP or reported any one of the following factors that may indicate an increased risk for HIV: in the past year, having unprotected sex with more than one person with unknown (or positive) HIV status, testing positive for a sexually transmitted infection (STI) (syphilis, gonorrhea, or chlamydia), or using injection drugs.

Data collection

The institutional review boards of the affiliated hospitals approved this study prior to data collection. An employee at each FQHC acted as a study contact and assisted with recruitment. The contacts advertised the study through word-of-mouth to coworkers and relayed the contact information of those interested to research staff. Patients were informed about the study from FQHC employees and flyers while visiting the FQHC for HIV testing. Those interested filled out consent-to-contact forms, which were securely and electronically sent to research staff. Potential participants were then contacted by a research assistant, screened for eligibility, electronically consented via DocuSign (a HIPAA-compliant signature capturing program), then scheduled for an interview. Interviews occurred remotely over Zoom, a HIPAA-compliant, video conferencing platform. Interviews were conducted until data saturation was reached. In addition to the interview, all participants were asked to complete a short demographics survey via REDCap, a HIPAA-compliant, online, data collection tool. Each participant received a $100 gift card for their time.

The i-PARIHS framework guided interview content and was used to create a semi-structured interview guide [ 28 ]. Within the i-PARIHS framework’s elements, the interview guide content included facilitators and barriers to PrEP use at the FQHC: 1) the innovation, (PrEP), such as its degree of fit with existing practices and values at FQHCs; 2) the recipients (individuals presenting to FQHCs), such as their PrEP awareness, barriers to receiving PrEP such as motivation, resources, support, and personal PrEP experiences; and 3) the context of the setting (FQHCs), such as clinic staff PrEP awareness, barriers providing PrEP services, and recommendations regarding PrEP care. Interviews specifically asked about the use of telemedicine, various methods for expanding PrEP knowledge for both patients and providers (e.g., social media, advertisements, community events/seminars), and location of services (e.g., mobile clinics, gyms, annual health checkups, health fairs). Staff and patients were asked the same interview questions. Data were reviewed and analyzed iteratively throughout data collection, and interview guides were adapted as needed.

Data analysis

Interviews were all audio-recorded, then transcribed by an outside, HIPAA-certified transcription company. Transcriptions were reviewed for accuracy by the research staff who conducted the interviews.

Seven members of the research team (TA, LW, KKG, AB, CSG, AL, LKB) independently coded the transcripts using an a priori coding schedule that was developed using the i-PARIHS and previous studies [ 15 , 16 , 17 ]. All research team members were trained in qualitative methods prior to beginning the coding process. The coding scheme covered: patient PrEP awareness, clinic staff PrEP awareness, barriers to receiving PrEP services, barriers to providing PrEP services, and motivation to take PrEP. Each coder read each line of text and identified if any of the codes from the a priori coding framework were potentially at play in each piece of text. Double coding was permitted when applicable. New codes were created and defined when a piece of text from transcripts represented a new important idea. Codes were categorized according to alignment with i-PARIHS constructs. To ensure intercoder reliability, the first 50% of the interviews were coded by two researchers. Team meetings were regularly held to discuss coding discrepancies (to reach a consensus). Coded data were organized using NVivo software (Version 12). Data were deductively analyzed using reflexive thematic analysis, a six-step process for analyzing and reporting qualitative data, to determine themes relevant to selecting appropriate implementation strategies to increase PrEP use at FQHCs in MS [ 29 ]. The resulting thematic categories were used to select ERIC implementation strategies [ 26 ]. Elements for each strategy were then operationalized and the mechanism of change for each strategy was hypothesized [ 30 , 31 ]. Mechanisms define how an implementation strategy will have an effect [ 30 , 31 ]. We used the identified determinants to hypothesize the mechanism of change for each strategy.

Member checking focus groups

Member checking is when the data or results are presented back to the participants, who provide feedback [ 32 ] to check for accuracy [ 33 ] and improve the validity of the data [ 34 ]. This process helps reduce the possibility of misrepresentation of the data [ 35 ]. Member checking was completed with clinic staff rather than patients because the focus was on identifying strategies to implement PrEP in the FQHCs.

Two focus groups were conducted with nine staff from the three FQHCs in MS. Eligibility criteria were the same as above. A combination of previously interviewed staff and non-interviewed staff were recruited. Staff members were a mix of medical (e.g., nurses, patient navigators, social workers) and non-medical (e.g., administrative assistant, branding officer) personnel. Focus group one had six participants and focus group two had three participants. The goal was for focus group participants to comprise half of staff members who had previously been interviewed and half of non-interviewed staff.

Participants were recruited and compensated via the same methods as above. All participants electronically consented via DocuSign, and then were scheduled for a focus group. Focus groups occurred remotely over Zoom. Focus groups were conducted until data saturation was reached and no new information surfaced. The goal of the focus groups was to member-check results from the interviews and assess the feasibility and acceptability of selected implementation strategies. PowerPoint slides with the results and implementation strategies written in lay terms were shared with the participants, which is a suggested technique to use in member checking [ 33 ]. Participants were asked to provide feedback on each slide.

Focus groups were all audio-recorded, then transcribed. Transcriptions were reviewed for accuracy by the research staff who completed focus groups. Findings from the focus groups were synthesized using rapid qualitative analyses [ 36 , 37 ]. Facilitators (TA, PPE) both took notes during the focus groups of the primary findings. Notes were then compared during team meetings and results were finalized. Results obtained from previous findings of the interviews and i-PARIHS framework were presented. To ensure the reliability of results, an additional team member (KKG) read the transcripts to verify the primary findings and selected supportive quotes for each theme. Team meetings were regularly held to discuss the results.

Thirty-six semi-structured interviews in HIV hot spots were completed between April 2021 and March 2022. Among the 19 FQHC staff, most staff members had several years of experience working with those at risk for HIV. Staff members were a mix of medical (e.g., doctors, nurses, CNAs, social workers) and non-medical (e.g., receptionists, case managers) personnel. Table 1 provides the demographic characteristics for the 19 FQHC clinic staff and 17 FQHC patients.

Table 2 provides a detailed description of the findings within each category: PrEP knowledge, PrEP barriers, and PrEP motivation. Themes are described in detail, with representative quotes, below. Implementation determinants are specific factors that influence implementation outcomes and can be barriers or facilitators. Table 3 highlights which implementation determinants can increase ( +) or decrease (-) the implementation of PrEP at FQHCs in MS. Each determinant, mapped to its corresponding i-PARIHS construct, is discussed in more detail below. There were no significant differences in responses across the three FQHCs.

PrEP knowledge

Patient prep awareness (i-parihs: recipients).

Most patients had heard of PrEP and were somewhat familiar with the medication. One patient described her knowledge of PrEP as follows, “I know that PrEP is I guess a program that helps people who are high-risk with sexual behaviors and that doesn't have HIV, but they're at high-risk.”- Patient, Age 32, Female, Not on PrEP. However, many lacked knowledge of who may benefit from PrEP, where to receive a prescription, the different medications used for PrEP, and the efficacy of PrEP. Below is a comment made by a patient listing what she would need to know to consider taking PrEP. “I would need to know the price. I would need to know the side effects. I need to know the percentage, like, is it 100 or 90 percent effective.”— Patient, Age Unknown, Female, Not on PrEP. Patients reported learning about PrEP via television and social media commercials, medical providers, and their social networks. One patient reported learning about PrEP from her cousin. “The only person I heard it [PrEP] from was my cousin, and she talks about it all the time, givin’ us advice and lettin’ us know that it’s a good thing.”— Patient, Age Unknown, Female, Not on PrEP.

Clinic Staff PrEP Awareness (i-PARIHS: Context)

Training in who may benefit from PrEP and how to prescribe PrEP varied among clinic staff at different FQHCs. Not all clinics offered formal PrEP education for employees; however, most knew that PrEP is a tool used for HIV prevention. Staff reported learning about PrEP via different speakers and meetings. A clinic staff member reported learning about PrEP during quarterly meetings. “Well, sometimes when we have different staff meetings, we have them quarterly, and we discuss PrEP. Throughout those meetings, they tell us a little bit of information about it, so that's how I know about PrEP.” – Staff, Dental Assistant, Female. Some FQHC staff members reported having very little knowledge of PrEP. One staff member shared that she knew only the “bare minimum” about PrEP, stating,

“I probably know the bare minimum about PrEP. I know a little about it [PrEP] as far as if taken the correct way, it can prevent you from gettin’ HIV. I know it [PrEP] doesn’t prevent against STDs but I know it’s a prevention method for HIV and just a healthier lifestyle.” –Staff, Accountant, Female

A few of the organizations had PrEP navigators to which providers refer patients. These providers were well informed on who to screen for PrEP eligibility and the process for helping the patient obtain a PrEP prescription. One clinic staff member highlighted how providers must be willing to be trained in the process of prescribing PrEP and make time for patients who may benefit. Specifically, she said,

“I have been trained [for PrEP/HIV care]. It just depends on if that’s something that you’re willing to do, they can train on what labs and stuff to order ’cause it’s a whole lot of labs. But usually, I try to do it. At least for everybody that’s high-risk.” – Staff, OB/GYN Nurse Practitioner, Female

Another clinic staff member reported learning about PrEP while observing another staff member being training in PrEP procedures.

“Well, they kinda explained to me what it [PrEP] is, but I was in training with the actual PrEP person, so it was kinda more so for his training. I know what PrEP is. I know the medications and I know he does a patient assistance program. If my patients have partners who are not HIV positive and wanna continue to be HIV negative, I can refer 'em.” – Staff, Administrative Assistant, Female

PrEP barriers

Barriers receiving prep services (i-parihs: recipients, innovation).

Several barriers to receiving PrEP services were identified in both patient and clinic staff interviews. There was a strong concern for the side effects of PrEP. One patient heard that PrEP could cause weight gain and nightmares, “I’m afraid of gaining weight. I’ve heard that actual HIV medication, a lotta people have nightmares or bad dreams.” - Patient, Age 30, Female, Not on PrEP. Another patient was concerned about perceived general side effects that many medications have. “Probably just the [potential] side effects. You know, most of the pills have allergic reactions and side effects, dizziness, seizures, you know.” - Patient, Age 30, Female, Not on PrEP.

The burden of remembering to take a daily pill was also mentioned as a barrier to PrEP use. One female patient explained how PrEP is something she is interested in taking; however, she would be unable to take a daily medication.

“I’m in school now and not used to takin’ a medication every day. I was takin’ a birth control pill, but now take a shot. That was one of the main reasons that I didn’t start PrEP cause they did tell me I could get it that day. So like I wanna be in the mind state to where I’m able to mentally, in my head, take a pill every day. PrEP is somethin’ that I wanna do.” - Patient, Age Unknown, Female, Not on PrEP

Stigma and confidentiality were also barriers to PrEP use at FQHCs. One staff member highlighted how in small communities it is difficult to go to a clinic where employees know you personally. Saying,

“If somebody knows you’re going to talk to this specific person, they know what you’re goin’ back there for, and that could cause you to be a little hesitant in coming. So there’s always gonna be a little hesitancy or mistrust, especially in a small community. Everybody knows everybody. The people that you’re gonna see goes to church with you.” – Staff, Accountant, Female

Some patients had a low perceived risk of HIV and felt PrEP may be an unnecessary addition to their routine. One patient shared that if she perceived she was at risk for HIV, then she would be more interested in taking PrEP, “If it ever came up to the point where I would need it [PrEP], then yes, I would want to know more about it [PrEP].”— Patient, Age Unknown, Female, Not on PrEP.

Some participants expressed difficulty initiating or staying on PrEP because of associated costs, transportation and/or scheduling barriers. A staff member explained how transportation may be available in the city but not available in more rural areas,

“I guess it all depends on the person and where they are. In a city it might take a while, but at least they have the transportation compared to someone that lives in a rural area where transportation might be an issue.” - Staff, Director of Nurses, Female

Childcare during appointments was also mentioned as a barrier, “It looks like here a lot of people don't have transportation or reliable transportation and another thing I don't have anybody to watch my kids right now. —Staff, Patient Navigator, Female.

Barriers Providing PrEP Services (i-PARIHS: Context)

Barriers to providing PrEP services were also identified. Many providers are still not trained in PrEP procedures nor feel comfortable discussing or prescribing PrEP to their patients. One patient shared an experience of going to a provider who was PrEP-uninformed and assumed his medication was to treat HIV,

“Once I told her about it [PrEP], she [clinic provider] literally right in front of me, Googled it [PrEP], and then she was Googlin’ the medication, Descovy. I went to get a lab work, and she came back and was like, “Is this for treatment?” I was like, “Why would you automatically think it’s for treatment?” I literally told her and the nurse, “I would never come here if I lived here.” - Patient, Age 50, Male, Taking PrEP

Also, it was reported that there is not enough variety in the kind of providers who offer PrEP (e.g., OB/GYN, primary care). Many providers such as OB/GYNs could serve as a great way to reach individuals who may benefit from PrEP; however, patients reported a lack of PrEP being discussed in annual visits. “My previous ones (OB/GYN), they’ve talked about birth control and every other method and they asked me if I wanted to get tested for HIV and any STIs, but the conversation never came up about PrEP.” -Patient, Age Unknown, Female, Not on PrEP.

PrEP motivation

Motivation to take prep (i-parihs: recipients).

Participants mentioned several motivators that enhanced patient willingness to use PrEP. Many patients reported being motivated to use PrEP to protect themselves and their partners from HIV. Additionally, participants reported wanting to take PrEP to help their community. One patient reported being motivated by both his sexuality and the rates of HIV in his area, saying, “I mean, I'm bisexual. So, you know, anyway I can protect myself. You know, it's just bein' that the HIV number has risen. You know, that's scary. So just being, in, an area with higher incidents of cases.”— Patient, Age Unknown, Male, Not on PrEP . Some participants reported that experiencing an HIV scare also motivated them to consider using PrEP. One patient acknowledged his behaviors that put him at risk and indicated that this increased his willingness to take PrEP, “I was havin' a problem with, you know, uh, bein' promiscuous. You know? So it [PrEP] was, uh, something that I would think, would help me, if I wasn't gonna change the way I was, uh, actin' sexually.”— Patient, Age Unknown, Male, Taking PrEP .

Table 3 outlines the implementation strategies identified from themes from the interview and focus group data. Below we recognize the barriers and determinants to PrEP uptake for patients attending FQHCs in MS by each i-PARIHS construct (innovation, recipient, context) [ 28 ]. Based on the data, we mapped the determinants to specific strategies from the ERIC project [ 26 ] and hypothesized the mechanism of change for each strategy [ 30 , 31 ].

Two focus groups were conducted with nine staff from threeFQHCs in MS. There were six participants in the 1st focus group and three in the 2nd. Staff members were a mix of medical (e.g., nurses, patient navigators, social workers) and non-medical (e.g., administrative assistant, branding officer) personnel. Table 4 provides the demographic characteristics for the FQHC focus group participants.

Staff participating in the focus groups generally agreed that the strategies identified via the interviews were appropriate and acceptable. Focus group content helped to further clarify some of the selected strategies. Below we highlight findings by each strategy domain.

PrEP information dissemination

Participants specified that awareness of HIV is lower, and stigma related to PrEP is higher in rural areas. One participant specifically said,

“There is some awareness but needs to be more awareness, especially to rural areas here in Mississippi. If you live in the major metropolitan areas there is a lot of information but when we start looking at the rural communities, there is not a lot.” – Staff, Branding Officer, Male

Participants strongly agreed that many patients don’t realize they may benefit from PrEP and that more inclusive advertisements are needed. A nurse specifically stated,

“ When we have new clients that come in that we are trying to inform them about PrEP and I have asked them if they may have seen the commercial, especially the younger population. They will say exactly what you said, that “Oh, I thought that was for homosexuals or whatever,” and I am saying “No, it is for anyone that is at risk.” – Staff, Nurse, Female

Further, staff agreed that younger populations should be included in PrEP efforts to alleviate stigma. Participants added that including PrEP information with other prevention methods (i.e., birth control, vaccines) is a good place to include parents and adolescents:

“Just trying to educate them about Hepatitis and things of that nature, Herpes. I think we should also, as they are approaching 15, the same way we educate them about their cycle coming on and what to expect, it’s almost like we need to start incorporating this (PrEP education), even with different forms of birth control methods with our young ladies.” – Staff, Nurse, Female

Participants agreed that PrEP testimonials would be helpful, specifically from people who started PrEP, stopped, and then were diagnosed with HIV. Participants indicated that this may improve PrEP uptake and persistence. One nurse stated:

“I have seen where a patient has been on PrEP a time or two and at some point, early in the year or later part of the year, and we have seen where they’ve missed those appointments and were not consistent with their medication regimen. And we have seen those who’ve tested positive for HIV. So, if there is a way we could get one of those patients who will be willing to share their testimony, I think they can really be impactful because it’s showing that taking up preventive measures was good and then kind of being inconsistent, this is what the outcome is, unfortunately.” – Staff, Nurse, Female

Increase variety and number of PrEP providers

Participants agreed that a “PrEP champion” (someone to promote PrEP and answer PrEP related questions) would be helpful, especially for providers who need more education about PrEP to feel comfortable prescribing. A patient navigator said,

“I definitely think that a provider PrEP champion is needed in every clinic or organization that is offering PrEP. And it goes back to what we were saying about the providers not being knowledgeable on it [PrEP]. If you have a PrEP champion that already knows this information, it is gonna benefit everybody, patients, patient advocates, the provider, everyone all around. Everyone needs a champion." – Staff, Patient Navigator, Female

Staff noted that they have walk-in appointments for PrEP available; however, they often have too many walk-in appointments to see everyone. They noted that having more resources and providers may alleviate this barrier for some patients:

“We still have challenges with people walking in versus scheduling an appointment, but we do have same day appointments. It is just hard sometimes because the volume that we have at our clinic and the number of patients that we have that walk in on a daily basis.” – Staff, Social Worker, Female

Enhance PrEP provider alliance and trust

Participants agreed that educational meetings would be beneficial and highlighted that meetings should happen regularly and emphasized a preference for in-person meetings. This is emphasized by the statement below,

“They should be in-person with handouts. You have to kind of meet people where they are as far as learning. Giving the knowledge, obtaining the knowledge, and using it, and so you have to find a place. I definitely think that yearly in-person training to update guidelines, medication doses, different things like that." – Staff, Patient Navigator, Female

Staff also suggested hosting one very large collaborative event to bring together all organizations that offer PrEP and HIV testing to meet and discuss additional efforts:

“What I would like to see happen here in the state of Mississippi, because we are so high on the list for new HIV infections, I would like to see a big collaborative event. As far as PrEP goes, those that are not on PrEP, one big collaborative event with different community health centers. You do testing, we do PrEP, and the referral get split. Everyone coming together for one main purpose.” – Staff, Patient Navigator, Female

Increase access to PrEP

Participants highlighted that most of the clinics they worked for already offer a variety of service sites (pharmacy, mobile clinic) but that more clinics should offer these alternative options for patients to receive PrEP. One patient navigator outlined the services they offer,

“We have a mobile unit. We do not have a home health travel nurse. We do telephone visits. We offer primary care, OB/GYN. We have our own pharmacy. We also have samples in our pharmacy available to patients that can’t get their medicine on the same day cos we like to implement same day PrEP. It has worked for us. More people should utilize those services.” – Staff, Patient Navigator, Female

Other staff suggested utilizing minute clinics and pharmacies at grocery stores. Highlighting, that offering PrEP at these locations may increase PrEP uptake.

There has been great scientific expansion of HIV prevention research and priorities must now pivot to addressing how to best implement effective interventions like PrEP [ 38 ]. PrEP remains underutilized among individuals who may benefit, particularly in Southern states such as MS [ 10 , 11 , 12 , 13 , 14 ]. Implementation science could help ameliorate this by identifying barriers and facilitators to PrEP rollout and uptake. We selected and defined several strategies from the ERIC project [ 26 ] to increase PrEP use utilizing FQHCs. Our results, as shown in Table  3 , highlight the four domains of strategies selected: 1) PrEP Information Dissemination, 2) Increase Variety and Number of PrEP Providers, 3) Enhance PrEP Provider Alliance and Trust, and 4) Increase Access to PrEP.

Firstly, individuals cannot utilize PrEP if they are not aware of its presence and utility. In Mississippi, advertising PrEP services is integral to implementation efforts given the existing stigma and lack of health literacy in this region [ 39 ]. Potential avenues for expanding PrEP awareness are integrating it into educational curriculums, adolescents’ routine preventative healthcare, and health fairs. This study compliments prior research that people should be offered sexual health and PrEP education at a younger age to increase awareness of risk, foster change in social norms and enhance willingness to seek out prevention services [ 40 , 41 ]. To meet the resulting growing need for PrEP educators, healthcare professionals should receive up-to-date PrEP information and training, so that they can confidently relay information to their patients. Similar to existing research, increasing provider education could accelerate PrEP expansion [ 42 , 43 , 44 ]. Training programs aimed at increasing provider PrEP knowledge may increase PrEP prescriptions provided [ 43 ] by addressing one of the most frequently listed barriers to PrEP prescription among providers [ 45 , 46 ].

Many patients prefer to receive PrEP at the healthcare locations they already attend and report a barrier to PrEP being limited healthcare settings that offer PrEP [ 39 , 47 , 48 , 49 ]. The aforementioned PrEP training could increase the number of healthcare workers willing to provide PrEP services. It is also imperative that providers in a diverse range of healthcare settings (e.g., primary care, OB/GYN, pediatricians and adolescent medicine providers) join the list of those offering PrEP to reduce stigma and enhance patient comfort.

These results mirrored other studies in the South that have shown that using relatable healthcare providers and trusted members of the community may serve to facilitate PrEP uptake [ 41 , 50 , 51 ]. If patients have a larger number of PrEP providers to choose from, they can select one that best fits their needs (e.g., location, in-network) and preferences (e.g., familiarity, cultural similarities). Enhanced comfort facilitates a strong patient-provider alliance and can lead to more open/honest communication regarding HIV risk behavior.

The lack of conveniently located PrEP providers is consistently reported as a structural barrier in the South [ 44 , 52 ]. This creates an increase in the demand on patients to attend regular follow-up appointments. The three strategies above all play a vital role in increasing access to PrEP. If more individuals are trained to provide PrEP care, there will be more PrEP providers, and patients can choose the best option for them. A sizeable influx of new PrEP providers could help staff new care facilities and service options in the community (e.g., mobile health units, home care, community-based clinics, telemedicine). Offering PrEP via telemedicine and mobile clinics to patients has been largely supported in the literature [ 44 , 53 , 54 ]. Intra- and inter-organizational collaborations could similarly increase PrEP access by sharing information and resources to ensure patients get timely, reliable care.

Our results largely supported previous findings by two systematic reviews on the barriers to PrEP uptake and implementation strategies to overcome it [ 39 , 47 ]. Sullivan et.al.’s review focused on the Southern U.S. [ 38 ], while Bonacci et. al. explored steps to improve PrEP equity for Black and Hispanic/Latino communities [ 47 ]. Both agreed that barriers to PrEP access are complex. Thus, cooperation from policymakers and the expansion of state Medicaid or targeted Medicaid waivers is vital to make PrEP attainable for those living in the coverage gap. Further, many FQHCs receive Ryan White funding for HIV care and treatment, contracting flexibility in the utility of these other sources of support may aid in eliminating the cost of PrEP as a barrier. They also stressed the need for educating community members and healthcare personnel about PrEP, increasing and diversifying PrEP service sites, normalizing PrEP campaigns and screening to alleviate stigma, and streamlining clinical procedures to facilitate the option for same-day PrEP. However, they also noted that these strategies are easier said than done. This further highlights the need for prioritizing research efforts towards implementation studies for effectiveness and practicality of overcoming the complex and systemic needs around HIV prevention/treatment.

The present study was able to build on past findings by providing a more holistic view of the barriers to PrEP use and possible strategies to address them through querying PrEP-eligible patients, medical providers, and non-medical staff. By interviewing a diverse range of stakeholders, it was possible to identify unmet patient needs, current PrEP care procedures and infrastructure, and attitudes and needed resources among those who could potentially be trained to provide PrEP in the future.

Limitations

Our results are limited to participants and clinic staff who were willing to engage in a research interview to discuss PrEP and FQHCs. Results are only generalizable to Mississippi and may be less relevant for other geographic areas. However, this is a strength given these strategies are meant to be tailored specifically to FQHCs in MS. Due to COVID-19 restrictions, interviews were conducted via Zoom. This allowed us to reach participants unable to come in physically for an interview and may have increased their comfort responding to questions [ 55 ]. However, some participants may have been less comfortable discussing via Zoom, which may have limited their willingness to respond.

This study highlighted the need for implementing PrEP strategies to combat HIV in Mississippi. PrEP knowledge, barriers, and motivation were identified as key factors influencing PrEP utilization, and four domains of strategies were identified for improving PrEP accessibility and uptake. Future research should further refine and assess the feasibility and acceptability of selected and defined implementation strategies and test strategies.

Availability of data and materials

De-identified data from this study are not available in a public archive due to sensitive nature of the data. De-identified data from this study will be made available (as allowable according to institutional IRB standards) by emailing the corresponding author.

Abbreviations

Mississippi

Pre-Exposure Prophylaxis

Federally Qualified Health Centers

Integrated-Promoting Action on Research Implementation in Health Services

Expert Recommendations for Implementing Change

Men Who Have Sex With Men

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Acknowledgements

Authors would like to acknowledge and thank Sarah Bailey for reviewing the manuscript and assisting for formatting.

This study was funded by the National Institute of Health (R34MH115744) and was facilitated by the Providence/Boston Center for AIDS Research (P30AI042853). Additionally, work by Dr. Trisha Arnold was supported by the National Institute of Mental Health Grant (K23MH124539-01A1) and work by Dr. Andrew Barnett was supported by the National Institute of Mental Health Grant (T32MH078788). Dr. Elwy is supported by a Department of Veterans Affairs Research Career Scientist Award (RCS 23–018).

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Trisha Arnold, Kayla K. Giorlando, Andrew P. Barnett, Avery Leigland & Larry K. Brown

Department of Psychiatry and Human Behavior, Warren Alpert Medical School of Brown University, Providence, RI, USA

Trisha Arnold, Laura Whiteley, Andrew P. Barnett, Ariana M. Albanese, A. Rani Elwy & Larry K. Brown

Department of Population Health Science, University of Mississippi Medical Center, Jackson, MS, USA

Courtney Sims-Gomillia & Precious Patrick Edet

Department of Medicine, University of Mississippi Medical Center, Jackson, MS, USA

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Contributions

TA and ARE led the conceptualization of this paper. TA, LW, LKB, DML, and JBB completed the literature search and study design. TA, LW, LKB, KKG, PPE, AB, AL, and CSG assisted with analyzing and interpreting the data. TA, ARE, and AMA finalized the results and implementation concepts of the study. All authors read and approved the final manuscript.

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Correspondence to Trisha Arnold .

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Arnold, T., Whiteley, L., Giorlando, K.K. et al. A qualitative study identifying implementation strategies using the i-PARIHS framework to increase access to pre-exposure prophylaxis at federally qualified health centers in Mississippi. Implement Sci Commun 5 , 92 (2024). https://doi.org/10.1186/s43058-024-00632-6

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Writing a Personal Statement

Preparing to Write

Brainstorming, don't forget, sample prompts.

A personal statement is a narrative essay that connects your background, experiences, and goals to the mission, requirements, and desired outcomes of the specific opportunity you are seeking. It is a critical component in the selection process, whether the essay is for a competitive internship, a graduate fellowship, or admittance to a graduate school program. It gives the selection committee the best opportunity to get to know you, how you think and make decisions, ways in which past experiences have been significant or formative, and how you envision your future. Personal statements can be varied in form; some are given a specific prompt, while others are less structured. However, in general a personal statement should answer the following questions:

• Who are you?
• What are your goals?
• How does this specific program/opportunity help you achieve your goals?
• What is in the future?

A personal statement is not:

• A variation of your college admissions essay
• An academic/research paper
• A narrative version of your resume
• A creative writing piece (it can be creative, though)
• An essay about somebody else

Keep in mind that your statement is only a portion of the application and should be written with this in mind. Your entire application package will include some, possibly all, of the materials listed below. You will want to consider what these pieces of the application communicate about you. Your personal statement should aim to tie everything together and fill in or address any gaps. There will likely be some overlap but be sure not to be too repetitive.

• Personal Statement(s)
• Transcripts
• Letters of recommendations
• Sample of written work
• Research proposal

For a quick overview of personal statements, you might begin by watching this "5 Minute Fellowships" video!

If you are writing your first personal statement or working to improve upon an existing personal statement, the video below is a helpful, in-depth resource.

A large portion of your work towards completing a personal statement begins well before your first draft or even an outline. It is incredibly important to be sure you understand all of the rules and regulations around the statement. Things to consider before you begin writing:

• How many prompts? And what are they? It is important to know the basics so you can get your ideas in order. Some programs will require a general statement of interest and a focused supplementary or secondary statement closely aligned with the institution's goals.
• Are there formatting guidelines? Single or double spaced, margins, fonts, text sizes, etc. Our general guideline is to keep it simple.
• How do I submit my statement(s)? If uploading a document we highly suggest using a PDF as it will minimize the chances of accidental changes to formatting. Some programs may event ask you to copy and paste into a text box.
• When do I have to submit my statement(s)? Most are due at the time of application but some programs, especially medical schools, will ask for secondary statements a few months after you apply. In these instances be sure to complete them within two weeks, any longer is an indication that you aren't that interested in the institution.

Below is a second 5 Minute Fellowships video that can help you get started!

Before you start writing, take some time to reflect on your experiences and motivations as they relate to the programs to which you are applying. This will offer you a chance to organize your thoughts which will make the writing process much easier. Below are a list of questions to help you get started:

• What individuals, experiences or events have shaped your interest in this particular field?
• What has influenced your decision to apply to graduate school?
• How does this field align with your interests, strengths, and values?
• What distinguishes you from other applicants?
• What would you bring to this program/profession?
• What has prepared you for graduate study in this field? Consider your classes at Wellesley, research and work experience, including internships, summer jobs and volunteer work.
• Why are you interested in this particular institution or degree program?
• How is this program distinct from others?
• What do you hope to gain?
• What is motivating you to seek an advanced degree now?
• Where do you see yourself headed and how will this degree program help you get there?

For those applying to Medical School, if you need a committee letter for your application and are using the Medical Professions Advisory Committee you have already done a lot of heavy lifting through the 2017-2018 Applicant Information Form . Even if you aren't using MPAC the applicant information form is a great place to start.

Another great place to start is through talking out your ideas. You have a number of options both on and off campus, such as: Career Education advisors and mentors ( you can set up an appointment here ), major advisor, family, friends. If you are applying to a graduate program it is especially important to talk with a faculty member in the field. Remember to take good notes so you can refer to them later.

When you begin writing keep in mind that your essay is one of many in the application pool. This is not to say you should exaggerate your experiences to “stand out” but that you should focus on clear, concise writing. Also keep in mind that the readers are considering you not just as a potential student but a future colleague. Be sure to show them examples and experiences which demonstrate you are ready to begin their program.

It is important to remember that your personal statement will take time and energy to complete, so plan accordingly. Every application and statement should be seen as different from one another, even if they are all the same type of program. Each institution may teach you the same material but their delivery or focus will be slightly different.

In addition, remember:

• Be yourself: You aren’t good at being someone else
• Tragedy is not a requirement, reflection and depth are
• Research the institution or organization
• Proofread, proofread, proofread
• How to have your personal statement reviewed

The prompts below are from actual applications to a several types of programs. As you will notice many of them are VERY general in nature. This is why it is so important to do your research and reflect on your motivations. Although the prompts are similar in nature the resulting statements would be very different depending on the discipline and type of program, as well as your particular background and reasons for wanting to pursue this graduate degree.

• This statement should illustrate your academic background and experiences and explain why you would excel in the Department of Civil and Environmental Engineering (UMass Amherst - M.S. in Civil Engineering).
• Describe your academic and career objectives and how the Yale School of Forestry and Environmental Studies can help you achieve them. Include other considerations that explain why you seek admissions to the Yale School of Forestry and Environmental Studies and your interests in the environmental field (Yale - Master of Environmental Management).  
• Please discuss your academic interests and goals. Include your current professional and research interests, as well as your long-range professional objectives. Please be as specific as possible about how your objectives can be met at Clark and do not exceed 800 words (Clark University - M.A. in International Development and Social Change).
• Write a 500- to 700-word statement that describes your work or research. Discuss how you came to focus on the medium, body of work, or academic area you wish to pursue at the graduate level. Also discuss future directions or goals for your work, and describe how the Master of Fine Arts in Studio (Printmedia) is particularly suited to your professional goals (School of the Art Institute of Chicago - MFA in Studio, Printmaking).
• Your statement should explain why you want to study economics at the graduate level. The statement is particularly important if there is something unusual about your background and preparation that you would like us to know about you (University of Texas at Austin - Ph.D in Economics).
• Your personal goal statement is an important part of the review process for our faculty members as they consider your application. They want to know about your background, work experience, plans for graduate study and professional career, qualifications that make you a strong candidate for the program, and any other relevant information (Indiana University Bloomington - M.S.Ed. in Secondary Education).
• Your autobiographical essay/personal statement is a narrative that outlines significant experiences in your life, including childhood experiences, study and work, your strengths and aspirations in the field of architecture, and why you want to come to the University of Oregon (University of Oregon - Master of Architecture).
• Personal history and diversity statement, in which you describe how your personal background informs your decision to pursue a graduate degree. You may refer to any educational, familial, cultural, economic or social experiences, challenges, community service, outreach activities, residency and citizenship, first-generation college status, or opportunities relevant to your academic journey; how your life experiences contribute to the social, intellectual or cultural diversity within a campus community and your chosen field; or how you might serve educationally underrepresented and underserved segments of society with your graduate education (U.C. Davis - M.A. in Linguistics).
• A Personal Statement specifying your past experiences, reasons for applying, and your areas of interest. It should explain your intellectual and personal goals, why you are interested in pursuing an interdisciplinary degree rather than a more traditional disciplinary one, and how this degree fits into your intellectual and personal future (Rutgers University - Ph.D in Women’s and Gender Studies).
• Your application requires a written statement to uploaded into your application and is a critical component of your application for admission. This is your opportunity to tell us what excites you about the field of library and information science, and what problems you want to help solve in this field. Please also tell us how your prior experiences have prepared you for this next step toward your career goals and how this program will help you achieve them (University of North Carolina Chapel Hill - Master of Science in Library Science).
• After watching the video, please describe what strengths and preferences as a learner you have that will facilitate your success in this innovative curriculum. What challenges in our curriculum do you anticipate and what strategies might you use to address these challenges? (MGH Institute of Health Professions PT - They recently redesigned their curriculum)
• Your personal goal statement should briefly describe how you view the future of the field, what your goals are to be part of that future, and what brought you to pursue an advanced education degree in your chosen field. You may include any other information that you feel might be useful. (Northeastern PT)
• Personal Statement: In 500 words or less, describe a meaningful educational experience that affected your professional goals and growth and explain how it impacted you. The educational experience does not need to be related to this degree. Focus on the educational experience and not why you think you would be a good professional in this field. (Simmons PT)
• Personal Statement (500 word minimum): State your reasons for seeking admission to this program at this institution. Include your professional goals, why you want to pursue a career in this field and how admission to this program will assist you in accomplishing those goals. (Regis College Nursing)
• “Use the space provided to explain why you want to go to this type of program.” (AMCAS)
• Address the following three questions(Though there is no set limit, most statements are 1–2 pages, single-spaced.): What are your reasons for pursuing this degree? Why do you wish to pursue your degree at this institution? How do you intend to leverage your degree in a career of this field? (Boston University MPH)
• Please submit a personal statement/statement of purpose of no more than 500 words for the department/degree of choice. Professional degree essays require a clear understanding of the _______ field and how you hope to work within the field. Be sure to proofread your personal statement carefully for spelling and grammar. In your statement, be sure to address the following: what interests you in the field of _____ what interests you in a specific degree program and department at this institution and what interests you in a particular certificate (if applicable). Please also describe how you hope to use your ________ training to help you achieve your career goals. (Columbia PhD in Public Health - Epidemiology)
• Because each Home Program requires significant original research activities in fulfillment of the requirements for the degree, we are interested in obtaining as much information as possible about your previous research experiences. Those who already have such experience are in a better position to know whether they are truly interested in performing ______ research as part of a graduate program. Please include specific information about your research experience in your Statement of Purpose. You may also use the Statement to amplify your comments about your choice of Home Program(s), and how your past experiences and current interests are related to your choice. Personal Statements should not exceed two pages in length (single spaced). Make sure to set your computer to Western European or other English-language setting. We cannot guarantee the ability to access your statement if it is submitted in other fonts. (Stanford Biosciences PhD)
• Your statement of purpose should describe succinctly your reasons for applying to the Department of ____ at ___ University. It would be helpful to include what you have done to prepare for this degree program. Please describe your research interests, past research experience, future career plans and other details of your background and interests that will allow us to evaluate your ability to thrive in our program. If you have interests that align with a specific faculty member, you may state this in your application. Your statement of purpose should not exceed two pages in length (single spaced). (Stanford Bioengineering PhD)
• Statement of purpose (Up to one page or 1,000 words): Rather than a research proposal, you should provide a statement of purpose. Your statement should be written in English and explain your motivation for applying for the course at this institution and your relevant experience and education. Please provide an indication of the area of your proposed research and supervisor(s) in your statement. This will be assessed for the coherence of the statement; evidence of motivation for and understanding of the proposed area of study; the ability to present a reasoned case in English; and commitment to the subject. (Oxford Inorganic Chemistry - DPhil)

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• Associations between growth, maturation and injury in youth athletes engaged in elite pathways: a scoping review
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• http://orcid.org/0000-0002-1915-9324 Gemma N Parry 1 ,
• http://orcid.org/0000-0003-1460-0085 Sean Williams 1 ,
• Carly D McKay 1 ,
• David J Johnson 1 , 2 ,
• http://orcid.org/0000-0003-1377-0234 Michael F Bergeron 3 ,
• Sean P Cumming 1
• 1 Department of Health , University of Bath—Claverton Down Campus , Bath , UK
• 2 West Ham United Football Club , London , UK
• 3 Performance Health , WTA Women’s Tennis Association , St. Petersburg , Florida , USA
• Correspondence to Dr Gemma N Parry; gp799{at}bath.ac.uk

Objective To describe the evidence pertaining to associations between growth, maturation and injury in elite youth athletes.

Design Scoping review.

Data sources Electronic databases (SPORTDiscus, Embase, PubMed, MEDLINE and Web of Science) searched on 30 May 2023.

Eligibility criteria Original studies published since 2000 using quantitative or qualitative designs investigating associations between growth, maturation and injury in elite youth athletes.

Results From an initial 518 titles, 36 full-text articles were evaluated, of which 30 were eligible for final inclusion. Most studies were quantitative and employed prospective designs. Significant heterogeneity was evident across samples and in the operationalisation and measurement of growth, maturation and injury. Injury incidence and burden generally increased with maturity status, although growth-related injuries peaked during the adolescent growth spurt. More rapid growth in stature and of the lower limbs was associated with greater injury incidence and burden. While maturity timing did not show a clear or consistent association with injury, it may contribute to risk and burden due to variations in maturity status.

Conclusion Evidence suggests that the processes of growth and maturation contribute to injury risk and burden in elite youth athletes, although the nature of the association varies with injury type. More research investigating the main and interactive effects on growth and maturation on injury is warranted, especially in female athletes and across a greater diversity of sports.

• Athletic Injuries
• Sporting injuries

Data availability statement

All data relevant to the study are included in the article or uploaded as supplementary information. Data relevant to the study have been uploaded as supplementary information.

This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/ .

https://doi.org/10.1136/bjsports-2024-108233

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WHAT IS ALREADY KNOWN ON THIS TOPIC

Growth and maturation during adolescence have been identified as risk factors for potential injury in young athletes.

WHAT THIS STUDY ADDS

This review identified 30 contemporary studies. Injury incidence and burden appear most closely related to maturity status and tempo of growth, with growth-related injuries peaking during the adolescent growth spurt. Practitioners are advised to consider measures of growth and maturation alongside clinical and field-based measurements.

HOW THIS STUDY MIGHT AFFECT RESEARCH, PRACTICE OR POLICY

Several methodological limitations and inconsistencies exist in the current evidence. Greater consistency and agreement on measurement practices could benefit future research quality, with the inclusion of female and non-football populations to address large gaps in the literature.

Introduction

The relationship between growth, maturation and injury risk in youth athletes is a topic of increasing interest in sports medicine, with a particular focus on the adolescent growth spurt 1 . Initiated by changes in the endocrine system, adolescence is the transitional phase between childhood and adulthood, during which the body undergoes rapid changes in size, shape and composition. 2 It also involves transformation of the circulatory, respiratory and metabolic systems, resulting in substantial changes in athletic and functional capacity. 3 Although the processes of growth and maturation have been proposed as risk factors for injury in young athletes, the evidence to support this contention is limited. 4 Limitations within the research are, however, noted, including heterogeneity across samples, research designs and analytical methods, poor reporting quality and high loss to follow-up. 4

Associating growth and maturation with injury in youth athletes is a logical premise. 5 Physeal injuries are a unique consideration for those working with youth populations. 6 Rapid asynchronous growth in skeletal, muscular and ligamentous structures creates increased ligament stress transfer through relatively weaker physeal plates and bone layers, increasing risk for such injuries. Age-related changes in bone mineral density, imbalances between flexibility and strength and alterations in joint stiffness further contribute towards an increased susceptibility to fractures at the growth plate and apophyses. 5 7 Apophyseal and physeal injuries also follow a distal-to-proximal 8 gradient, consistent with the sequential and asynchronous nature of adolescent growth. 8 9 For example, Sever’s disease at the posterior calcaneus tends to present in advance of Osgood-Schlatter’s or Sinding-Larsen at the knee, which, in turn, present in advance of apophyseal injuries at the sites of the iliac crest and ischial tuberosity of the hip. Apophyseal injuries are attributed to the softening or malformation of the articular cartilage, the latter of which is associated with more rapid growth during adolescence. 7 From a perceptual-motor perspective, rapid and asynchronous changes in skeletal, muscular and ligamentous structures, coupled with developmental changes in neurocognitive processing, have also been linked to temporary disruptions in mobility and motor-coordination, which may further increase injury risk. 5

The introduction of injury surveillance and growth and maturation profiling systems in many elite performance pathways (eg, national athlete development programmes, professional sport academies) has afforded a more systematic and rigorous approach to monitoring physical development and health in young athletes. 10 Implemented in parallel and delivered by trained professionals and clinicians, these organised strategies have enabled the capture of high-quality longitudinal data in young athletes, stimulating further research related to growth, maturation and injury. Characterised by early specialisation and maintained elevated levels of training and competition, elite performance pathways may also provide a more conducive environment from which to observe and investigate associations between physical development and injury in youth. 1 Considering these advances, this scoping review endeavours to synthesise and expand on a previous systematic review, 4 with the aim of providing clinicians, researchers and other stakeholders with a description of contemporary research related to growth, maturation and injury in youth athletes engaged in current elite sports pathways, with a particular emphasis on the operationalisation of growth and maturation, methodological quality and assessment, sample populations, emerging evidence, knowledge gaps and limitations within the extant literature.

Equality diversity and inclusion statement

The author and article screening teams were gender-balanced and included senior and junior academic staff from multiple disciplines and professions. Articles were restricted to those published in English but were not excluded based on country of origin. Gender equity in the study of physical development and injury in young athletes is addressed in the discussion.

Information sources and search strategy

This review was commissioned by the International Olympic Committee to describe the extent, type and quality of contemporary research and evidence pertaining to growth, maturation and injury in ‘elite youth athletes’, contributing towards a consensus statement on the health, safety and sustainability of young athletes competing at the Olympic games. A definition of ‘elite youth athletes’ as ‘highly trained and invested youth athletes routinely participating at national level (Tier 3) to world-class (Tier 5) athletic competitions’ was established in advance of the review 11 12 and extended to include dancers enrolled in professional dance schools.

The search was conducted, in general, adhering to Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) Extension for Scoping Reviews, PRISMA-ScR ( online supplemental figure S1 ). All 20+2 PRISMA-ScR reporting items were met, except for protocol registration. Five databases (SPORTDiscus, Embase, PubMed, MEDLINE and Web of Science) were searched for relevant articles. As the review aimed to inform a contemporary consensus statement regarding the health and well-being of elite young athletes, the article search was restricted to peer-reviewed publications written in English from 1 January 2000 to 30 May 2023. A series of unique search terms were employed to identify relevant articles. The Boolean Operators “OR” and “AND” were used to broaden the search results, define the population of interest (ie, elite youth athletes), limit the intended outcomes of our search (ie, injury) and link the search terms. An asterisk (*) was applied to some keywords to search the database for all endings of the word or phrase (eg, matur*). The final search strategy being: (1) “elite” AND “youth” AND “athlete”, (2) AND “growth” OR “matur*” OR “pubert*”, (3) AND “Injur*” OR “medial epicondyle apophysitis” OR “Proximal Humeral Epiphysitis” OR “stress fracture” OR “growth plate fracture” OR “Pars” OR “Femoroacetabular impingement” OR “apophysitis” OR “Sever’s” OR “Osgood” OR “osteochondrosis” OR “osteochondritis dissecans” OR “spondylolysis”.

Supplemental material

Eligibility criteria and description of eligible studies.

Inclusion criteria were articles based on primary research, using original data and related to elite youth athletes. To meet the definition of ‘elite’ and be included in the review, the samples within each study had to be described in a manner that aligned with the criteria for routine participation in national (Tier 3) to world-class (Tier 5) competition. 12 For example, athletes who were members of professional sports academies or national development programmes were considered to have met these criteria. Studies using quantitative designs had to include measures of injury and growth and/or maturation. Qualitative studies investigating associations between growth, maturation and injury in elite youth athletes were eligible for inclusion. Review articles and case studies were excluded. Two independent reviewers (GNP, SC) completed the review process in May 2023. Each reviewer screened the articles, first at the level of title and abstract and then at the level of full text, to judge if the eligibility criteria were met. Disagreements were resolved through discussion and final consensus.

Data synthesis

To ensure consistency in the operationalisation of growth and maturation, the following definitions were adopted. 2 Growth was defined as rate of change in size of the body, its parts, or the proportions of various parts (eg, cm per annum, kg per annum). Maturation refers to the processes of progress towards the mature state occurring in multiple biological systems (eg, endocrine, sexual, skeletal and somatic) and was defined in terms of status, tempo and timing. Status denoted the stage of maturation attained at a specific time point, (eg, pre-peak height velocity (PHV), circa-PHV, post-PHV) whereas tempo described the rate at which maturation occurs. Timing was defined as the age at which maturational events (eg, PHV) occurred and/or the degree to which an athlete was advanced, on-time or delayed in maturation relative to age-specific and sex-specific standards. The data extracted from the eligible studies were summarised descriptively ( online supplemental table S1 ) and appraised with respect to methodological quality.

Study selection

The initial search retrieved 871 citations of which 518 remained after removing duplicates. Full texts of 37 articles were reviewed to determine eligibility, 30 of which were eligible for final inclusion. The eligible articles included 28 quantitative and 2 qualitative studies ( figure 1 ).

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Study selection process based on inclusion and exclusion criteria.

Data collection and risk of bias assessment/analysis of quality

Descriptive data were extracted from eligible studies, comprising study design, population, measures and operationalisation of growth and maturation, and injury definition and outcomes (eg, incidence, burden) ( online supplemental table S1 ). The methodological quality of the quantitative studies (n=28), as judged on the Downs and Black Scale, 13 ranged from 7 to 19 out of 32 points ( online supplemental table S2 ). The Downs and Black Scale is a 28-item checklist used to assess the methodological quality of both randomised and non-randomised studies, evaluating factors such as reporting, external validity, internal validity (bias and confounding) and statistical power. The median quality score of these articles was low (14/32), attributable to study design and the absence of explicit explanations regarding variables of interest and consideration of confounding factors. While some articles included large samples, the distribution of participants by maturity status and/or timing categories was typically non-normal, resulting in inadequate statistical power in several studies. Most studies focused on soccer (n=19) and were exclusive to male participants (n=24). Only four quantitative studies included male and female athletes, and none considered female-only cohorts. The two qualitative papers appraised using the Joanna Briggs Institute (JBI) critical appraisal checklist for qualitative research 14 15 both met 8 out of the 10 criteria, with each noting a failure to consider potential researcher influence on the study findings as a limitation. A breakdown of the individual item scores for each article using the Downs and Black and JBI checklists is available in online supplemental file 2 .

Maturity status and injury

17 quantitative studies 8 16–31 adopting prospective designs and two qualitative studies 32 33 investigated associations between maturity status and injury ( table 1 ). 16 of the quantitative studies 8 16–20 22–31 employed non-invasive estimates of somatic maturation with estimated age at PHV (EA PHV) (n=11) and percentage of predicted (%PAH) or attained adult height (% AAH) (n=5) the most common methods. One study used an estimate of sexual maturation to classify youth handball players as mature or immature. Wik and colleagues 30 employed estimates of both skeletal and somatic maturation in study of maturation and injury in male track and field athletes. Most studies categorised athletes into groups based on maturity status, however, five studies treated maturation as a continuous variable. 19 24 26 27 30 Samples sizes varied from n=21 24  to n=502. 17 The methodological quality of the quantitative studies ranged from poor 29 to fair. 34 Two qualitative studies 32 33 investigated coaches’ and practitioners’ perceptions of how maturational status was related to injury risk in male and female gymnasts. Both of these studies achieved JBI scores of 8 out of 10, indicating a high quality of qualitative research.

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Associations between maturational status and injury for all eligible studies

The maturational stage associated with the adolescent growth spurt (ie, circa-PHV) appears to yield the greatest increase in injury incidence per 1000 hours, in papers reporting competition, training and overall time loss. When broad definitions of injury were observed, circa-PHV status was associated with greater incidences of overall, 25 non-contact 19 20 and traumatic 28 29 injuries in academy soccer, and higher incidences of all-complaint injuries in both male and female elite youth gymnasts with the peak approximating 90% PAH. 24 While the evidence suggests a general trend towards increased injury incidence and burden with advancing maturation, 16 17 21 the nature of reported associations varied relative to injury type. 8 17 Circa-PHV and pre-PHV status were associated with a higher incidence or prevalence of injuries classified as growth-related in track and field, 34 handball 21 and soccer, 8 17 but not squash. 18 Both qualitative studies also identified the growth spurt as a maturational stage where incidence and burden for growth-related and lower back injuries was greater. 32 33 Moreover, growth-related injuries followed a distal-to-proximal gradient aligned with the sequential and asynchronous nature of adolescent growth. 8 Specifically, these injuries tended to present first in the more distal segments of the lower limbs, before presenting in the more proximal segments of the lower extremities, and finally the trunk and pelvis. Consistent with this pattern, a higher probability for non-contact lower extremity injuries was observed in U13–14 soccer players during the earlier phases of the adolescent growth spurt. 26 A greater number of years to EA PHV was, however, associated with an increased probability for traumatic and overuse injuries in male and female alpine skiers. 27 Mature athletes, or those advanced in maturity status (post-PHV), exhibited the lowest incidence and burden of growth-related injuries, 8 22 23 34 yet evidenced a higher incidence and burden for muscular, cartilaginous and ligamentous injuries in soccer, 8 23 but not in handball where these injuries were higher in immature athletes. 21 Maturity status was unrelated to patellar tendinopathy in male and female ballet dancers. 31

Associations between advancing maturity status and injury burden were observed in several studies. As with injury incidence, associations with burden varied by time-loss definitions and injury type. Six studies in soccer 16 17 19 20 22 23 that used various definitions of time loss reported an increase in injury burden with advancing maturity, with one study documenting peak burden for non-contact injuries at 95% PAH. 19 There were notably higher burdens of growth-related injuries in athletes during the pre and circa-PHV stages in soccer. 17 22 23 The post-PHV stage is associated with a reduced burden for growth-related injury burden, 17 22 but an increase in burden for non-contact 19 and muscle and joint-related injury, 22 although these findings are currently limited to male soccer players.

Maturity timing and injury

10 quantitative studies 20 22 28 35–41 using prospective designs ( table 2 ) investigated associations between maturity timing and injury. Six studies employed estimates of somatic maturation with EA PHV (n=4) 29 40 41 and %PAH 20 37 (n=2) the preferred methods. There was substantial heterogeneity in the criteria used to define early maturation, on-time maturation and late maturation across methods. Four studies 35 36 38 39 employed estimates of skeletal age via hand-wrist radiographs. To define early maturation, on-time maturation and late maturation, three studies 35 36 38 used the traditional criterion of ±1 years SA-CA (skeletal age-chronological age); whereas one study used ±0.5 years SA-CA. 39 Nine studies 20 22 28 35–39 41 included male athletes and one study included males and females. 40 Eight of the studies involved soccer, 20 22 29 35–39 with single studies in track and field 41 and alpine skiing. 40 Sample sizes ranged n=26 29  to n=233. 38 The methodological quality of the studies was generally low, ranging from poor 29 to fair. 39

Associations between maturational timing and injury for all relevant selected studies

Associations between the timing of maturity and injury were observed across several, 22 29 36 38 39 but not all 20 35 37 studies of male soccer players. The nature and direction of these associations did, however, vary depending on injury type and the age range of the sample. A closer inspection of the findings suggested that the impact of maturity status was more important than the timing of maturity itself when investigating injury susceptibility in young athletes. Consistent with this hypothesis, U14 and U13–U19 players categorised as early-maturing presented a higher incidence of injuries associated with more advanced stages of maturation, including reinjury, injury to ligaments, tendons and muscles and injuries to the groin, head or face, and overall injuries. 36 38 Early-maturing U14 players also reported higher burdens for time loss, muscle, hamstring and joint/ligament-related injuries. 39 In contrast, players categorised as ‘on-time’, presented higher incidence rates for injuries associated with the early-to-mid stages of adolescence, including lower limb apophyseal injuries, anterior inferior iliac spine avulsions, Osgood-Schlatter’s, Sever’s and knee-related injuries in on-time than early maturing. 36 38 In comparison to late-maturing players, on-time U14 players presented higher rates of incidence for, groin injuries, tendinopathies and moderate injuries, 36 and higher burdens for joint/ligament, knee, anterior inferior iliac spine, growth-related and time-loss injuries. 22 39 Late-maturing players presented higher incidence rates for osteochondrosis, thigh, growth-related and major injuries than early-maturing players, and a higher incidence of tendinopathies and osteochondral disorder of the knee than early and on-time players. 36 38 Late-maturing track and field athletes also presented a higher incidence for foot, ankle and lower limb injuries compared with their peers. 41 Despite late-maturing alpine skiers presenting a higher prevalence of traumatic and overuse injuries, respectively, these values did not differ statistically from early-maturing skiers. 40

Growth and maturity tempo and injury

Nine quantitative studies 19 23 25 27 34 42–45 adopting prospective designs ( table 3 ) and two qualitative studies 32 33 investigated associations between rate of growth and/or maturation and injury. Rate of change in stature (n=9) and leg length (n=5) were the most frequent growth measures. Four studies assessed growth tempo in body mass and mass-for-stature. Singular studies considered growth of the foot, 42 torso 30 and tempo of skeletal maturation. 34 Six studies 19 23 25 43–45 involved male soccer players with the remaining studies involving male track and field athletes, 34 and dancers 42 and alpine skiers 27 of both sexes. Samples sizes ranged from n=46 42  to n=378. 45 The methodological quality of the studies was generally low, ranging from poor 43 to fair. 34 45

Associations between growth and maturation indices and injury for all relevant selected studies

More rapid gains in stature were associated with a higher incidence and/or probability of overall, 25 43 45 non-contact 19 and overuse 44 injuries in male academy soccer players, and bone and growth plate injuries in male track and field athletes. 34 Greater growth in stature was, however, associated with a reduced probability for acute injuries in U13–15 academy soccer players 44 and overall injuries in male and female alpine skiers. 27 More rapid gains in stature during PHV was associated with a higher burden for overall, knee and growth-related injuries, but not muscle, joint/ligament, knee or ankle injuries in U14 academy soccer players. 22 A non-linear (inverted-U) association between growth in stature and burden for non-contact injuries was reported in U13–U16 male soccer players, with peak estimated injury burden occurring at 4.17 cm per year. 19 More rapid growth of the lower limbs was associated with a higher probability for overall injuries in alpine skiing 27 and track and field, 34 overuse 44 injuries in soccer, and bone and growth plate in track and field. 34 As with stature, a non-linear association was observed between growth rate of the lower limbs with peak estimated injury burden occurring at a growth rate of 5.27 cm per year. 19

Rate of growth in body mass did not differentiate between injured and non-injured alpine skiers 27 and was not associated with injury probability in track and field athletes 34 and U10–15 soccer players. 44 Greater seasonal gains in body mass were observed in injured vs non-injured Italian soccer players; however, growth in mass did not emerge as a predictor of injury in a subsequent regression model. 25 With respect to mass-for-stature, more rapid gains in body mass index (BMI) were observed between injured vs non-injured Dutch soccer players, 43 but not their Italian counterparts. 25 Rate of change in BMI was unrelated to overall injury risk injury in alpine skiing 27 and overall, gradual and sudden-onset, bone and growth plate injuries in track and field. 34

In both qualitative studies, 32 33 more rapid gains in stature and mass were perceived as risk factors for injury in young gymnasts, with specific reference to stress fractures, growth-related, shoulder and lower back injuries. The risk was also perceived to be higher when accompanied with high or sudden spikes in training and competition loads, and restricted eating.

Trunk growth was unrelated to probability for overall, gradual onset, sudden onset, bone or growth plate-related injuries in track and field athletes. 34 More rapid growth of the foot was, however, associated with a higher risk for lumbar and lower extremity injury risk in male and female dancers. 42 Finally, more rapid changes in skeletal maturation were associated with an increased risk of bone injuries in a track and field athletes, yet was unrelated to risk of gradual-onset and sudden-onset, growth plate and overall injuries. 34

This review described contemporary research and emerging evidence related to growth, maturation and injury in youth athletes engaged in elite performance pathways. A total of 30 articles were deemed eligible for inclusion in this review with the majority involving male soccer players. The disparity between sports reflects the popularity of soccer, as a sport, and the significant investment by professional clubs and national governing bodies in the monitoring of physical development and health of young players. 10 Implemented in parallel, injury surveillance and growth and maturity profiling systems have enabled researchers to collect high-quality longitudinal data, consider the implications of growth and maturation across diverse injury types and perform more sophisticated methods of analysis. Those investigating the associations between growth, maturation and injury in young athletes should look to soccer for examples of good practice (see Monasterio, 8 22 23 39 Rommers, 44 45 Materne 38 and Hall 17 ), such as the aggregation of data across age groups, clubs and/or multiple seasons and consideration of different injury types and interactional effects. Wik’s investigation of both maturity status and tempo as risk factors across multiple injury types in track and field also serves as a good example of innovative practice within this field. 34

The lack of studies involving and/or exclusive to female athletes is a particular concern. The eligible studies that considered physical development and injury in female athletes all had comparatively small sample sizes and, thus, it is difficult to draw firm conclusions from the evidence. Injury research related to male athletes does not always generalise to female athletes. 46 The sex differences in human anatomy and physiology that emerge during puberty may predispose male and female athletes to variable levels of injury risk. 7 Whereas some studies have found female athletes to be at greater or reduced risk for certain types of injury, 46 47 others suggest more comprehensive research and data are still required to effectively inform injury prevention and treatment strategies specific to female athletes. 48 There is a need for sport’s national governing bodies to prioritise and invest in research related to physical development and injury in female athletes. Care must, however, be taken in the design and implementation of growth and maturity profiling strategies for female athletes with particular sensitivity around the collection, communication and use of data. 49 Data pertaining to physical development should be used to understand and support the health and development of the athletes and not as a criteria for the selection and exclusion of athletes. 49 Clinicians and researchers should seek to identify those injuries that female and, to a lesser extent, male athletes may be more or less susceptible to at different stages of maturation and consider strategies for the early detection and mitigation of conditions such as relative energy deficiency RED-S which can compromise physical development and the immediate and long-term health of youth athletes. 49 Researchers should also investigate the extent to which the processes of growth and maturation contribute to the elevated risk of medial collateral ligament (MCL) and anterior cruciate ligament (ACL) injuries observed in female athletes during adolescence. 7

The evidence reviewed suggests that susceptibility to injury in youth athletes varies relative to maturity status. Adopting a broad definition of injury, incidence and burden generally increases with advancing maturation. The nature of the association between maturity status and injury does, however, vary relative to injury type. The adolescent growth spurt signalled a marked increase in the incidence and burden of growth-related injuries, confirming the beliefs and experiences of coaches and medical practitioners. 32 33 Growth-related injuries were most prevalent at the onset and during the growth spurt, with apophyseal, osteochondrosis and avulsion prominent in numerous sport contexts. 8 18 21 22 33 34 Growth-related injuries also followed a distal-to-proximal gradient consistent with the sequential and asynchronous nature of adolescent growth, before declining in both incidence and burden post-PHV. In contrast, the incidence of muscular, cartilaginous and ligamentous injuries generally increased with advancing maturity status and were most prevalent in athletes categorised as mature and/or post-PHV. Increased susceptibility for such injuries post-PHV has been attributed to several factors associated with growth and maturation, including disruptions in neuromuscular control, insufficient muscle capacity, imbalance in muscle and tendon growth, and increase in moments of inertia in athletes’ segments. 8 Despite much of the data within the eligible studies being normalised to exposure per 1000 hours, uncertainties remain regarding the standalone impact of maturational status on injury incidence and burden, given the variations in activity content (eg, modality, frequency, intensity, location) that may comprise an individual’s exposure hours.

The absence of a consistent pattern of association between timing and injury suggested that maturing in advance or delay of one’s peers (ie, timing) was not an inherent risk for injury. Rather, the extent to which timing impacts an athlete’s maturational status and/or proximity to PHV appeared to be of greater relevance. Compatible with this logic, early maturing athletes presented a higher incidence and burden for injuries associated with more advanced stages of maturity (eg, tendinopathies, groin strains, joint and ligament injuries, functional muscle disorders). 22 36 38 39 Conversely, on-time and late maturing reported a higher incidence and burden for injuries associated with the earlier stages of the growth spurt (eg, lower limb apophyseal injuries, osteochondrosis, AIIS, Osgood-Schlatter’s, Sever’s). 22 36 38 39 41 Due to inherent maturity selection biases that exist in sport, 3 50–52 late-maturing athletes were under-represented in several of the eligible studies 22 36 38 39 resulting in insufficient statistical power to make effective comparisons across groups. As late maturing athletes experience the growth spurt at an age when training intensities and volumes are typically higher, they may be more susceptible to injuries associated with growth or overloading. 53 Any elevated injury risk associated with later maturation may, however, be mitigated by a lower rate of growth at PHV. Future studies may need to aggregate data across teams, sport or consecutive seasons to test this hypothesis or focus on sports or activities where late developers are more likely to be represented (ie, gymnastics, ballet, diving).

More rapid gains in stature and length of the lower limbs were associated with a higher incidence of overuse, non-contact and growth-related injuries across several sports. Monasterio et al , also observed more rapid growth in stature to be associated with a higher burden for overall and growth-related injuries during PHV. 22 Johnson et al , did, however, report a non-linear associations between burden and rate of growth in stature and of the lower limbs, with peak burdens occurring post-PHV and at a rate of approximately 4-to-5 cm per year. 19 The delayed effect of injuries sustained during PHV on injury burden post-PHV may explain the discrepancy in these results. No clear or consistent associations were observed between rate of growth in mass and mass-for-stature with injury. Pubertal gains in mass and mass-for-stature may have greater injury implications for athletes participating sports that involve frequent landing, jumping, pivoting and high intensity accelerations and decelerations. 5 They may also have greater impact for risk in female athletes who experience larger gains in absolute and relative fat-mass, and corresponding smaller gains in absolute and relative strength during puberty. 2 50 Further research on the growth and injury in female athletes is needed with a particular focus on the preventative benefits of targeted functional movement and neuromuscular strength training interventions. 54–56

Intervention studies designed to mitigate growth-related injuries in young athletes are currently lacking. However, in the interim, growth and maturity profiling can provide valuable information pertaining to maturational status, timing and growth rate of individual athletes, enabling practitioners to identify athletes at heightened risk for specific injuries and adapt their training and conditioning accordingly. 1 For example, the Athletic Skills Model (ASM) describes a maturity-matching strategy whereby athletes identified as circa-PHV are assigned to training groups with an increased emphasis on movement competency, core and lower body strength, mobility, balance, coordination, coupled with reductions training load, intensity and movement repetition. 57 A recent intervention aligned to theASM principles resulted in marked reductions in non-contact injury incidence and burden among academy soccer players identified as circa-PHV and at-risk. 58 Although promising, further encompassing research is required to validate these findings.

Strengths and limitations

The inclusion of several contemporary studies conducted across multiple clubs or seasons was a strength of this review. 8 17 22 23 36 38 39 44 45 With comparatively large samples and more rigorous approaches to data capture, these studies enabled more comprehensive and detailed investigations of growth, maturation and injury. These studies also provided valuable insight as to the complex, multifaceted and dynamic nature of the relationship between physical development and injury in youth. The myriad of methodological limitations highlighted by Swain et al 4 persist, making it challenging to generalise findings or draw firm conclusions from the existing literature on this topic. Although the quality of the two qualitative studies was deemed excellent, the methodological quality of the quantitative studies was generally low, ranging from poor to fair. To improve research quality and robustness, there is a need to develop a contemporary consensus statement on standardised evidence-informed best practices for assessing and estimating growth and maturation in young athletes. This review was commissioned by the IOC as a wider contribution towards a consensus statement, and as such adopted a harmonised methodology. Researchers may wish to consider alternative methodologies in additional future reviews. Building on previews reviews and commentaries, 3 4 59–61 particular attention should be paid to the operationalisation of growth and maturation and the criteria used to determine maturity status, timing and tempo of growth. It is equally important to consider the rationale for measurement, analytical quality control, frequency of measurement, the education of key stakeholders and how data are communicated to athletes, parents/guardians and coaches. 49 The validity and reliability of the various methods used to estimate maturation should also be considered with a particular focus on non-invasive estimates of somatic maturation. 3 62 63

The evidence reviewed indicates that variability in growth and maturation plays a contributing role in the risk of injury among youth athletes engaged in high performance pathways. The associations involved are intricate and diverse, requiring further research to comprehend the precursors, mechanisms and contextual factors that may predispose athletes to a higher risk of specific injuries (eg, those related to motor competence, functional capacity, training/competition load and content). It is recommended that sport’s governing bodies simultaneously implement comprehensive injury surveillance and growth and maturity profiling systems for youth, along with educational content and a current consensus statement on best/standardised practice in the estimation and monitoring of growth and maturation. Such an approach has the potential to enhance our understanding of the connections between growth, maturation and injury and provide the empirical basis for the subsequent development and implementation of injury prevention programmes.

Ethics statements

Patient consent for publication.

Not applicable.

Ethics approval

Research was approved by The University of Bath ethics committee #0122-80.

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X @GemmaNParry, @statman_sean, @Dr_CMCKay, @DrMBergeron_01, @phd_Sean

Contributors GNP, SC and SW conceptualised and planned the study design. GNP and SC led the study. GNP and SC contributed to article screening. GNP is guarantor and accepts full responsibility for the work and/or the conduct of the study. GNP and SC has access to the data, and controlled the decision to publish. GNP, SC and SW wrote the first draft of the article. All coauthors reviewed, revised and approved the final manuscript.

Funding The research was funded via a consultancy award from the International Olympic Committee

Competing interests None declared.

Provenance and peer review Not commissioned; externally peer reviewed.

Supplemental material This content has been supplied by the author(s). It has not been vetted by BMJ Publishing Group Limited (BMJ) and may not have been peer-reviewed. Any opinions or recommendations discussed are solely those of the author(s) and are not endorsed by BMJ. BMJ disclaims all liability and responsibility arising from any reliance placed on the content. Where the content includes any translated material, BMJ does not warrant the accuracy and reliability of the translations (including but not limited to local regulations, clinical guidelines, terminology, drug names and drug dosages), and is not responsible for any error and/or omissions arising from translation and adaptation or otherwise.

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• Open access
• Published: 26 August 2024

Inter-laboratory comparison of eleven quantitative or digital PCR assays for detection of proviral bovine leukemia virus in blood samples

• Aneta Pluta 1 , 13 ,
• Juan Pablo Jaworski 2 ,
• Casey Droscha 3 ,
• Sophie VanderWeele 3 ,
• Tasia M. Taxis 4 ,
• Stephen Valas 5 ,
• Dragan Brnić 6 ,
• Andreja Jungić 6 ,
• María José Ruano 7 ,
• Azucena Sánchez 7 ,
• Kenji Murakami 8 ,
• Kurumi Nakamura 8 ,
• Rodrigo Puentes 9 ,
• MLaureana De Brun 9 ,
• Vanesa Ruiz 2 ,
• Marla Eliana Ladera Gómez 10 ,
• Pamela Lendez 10 ,
• Guillermina Dolcini 10 ,
• Marcelo Fernandes Camargos 11 ,
• Antônio Fonseca 11 ,
• Subarna Barua 12 ,
• Chengming Wang 12 ,
• Aleksandra Giza 13 &
• Jacek Kuźmak 1  

BMC Veterinary Research volume  20 , Article number:  381 ( 2024 ) Cite this article

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Bovine leukemia virus (BLV) is the etiological agent of enzootic bovine leukosis and causes a persistent infection that can leave cattle with no symptoms. Many countries have been able to successfully eradicate BLV through improved detection and management methods. However, with the increasing novel molecular detection methods there have been few efforts to standardize these results at global scale. This study aimed to determine the interlaboratory accuracy and agreement of 11 molecular tests in detecting BLV. Each qPCR/ddPCR method varied by target gene, primer design, DNA input and chemistries. DNA samples were extracted from blood of BLV-seropositive cattle and lyophilized to grant a better preservation during shipping to all participants around the globe. Twenty nine out of 44 samples were correctly identified by the 11 labs and all methods exhibited a diagnostic sensitivity between 74 and 100%. Agreement amongst different assays was linked to BLV copy numbers present in samples and the characteristics of each assay (i.e., BLV target sequence). Finally, the mean correlation value for all assays was within the range of strong correlation. This study highlights the importance of continuous need for standardization and harmonization amongst assays and the different participants. The results underscore the need of an international calibrator to estimate the efficiency (standard curve) of the different assays and improve quantitation accuracy. Additionally, this will inform future participants about the variability associated with emerging chemistries, methods, and technologies used to study BLV. Altogether, by improving tests performance worldwide it will positively aid in the eradication efforts.

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Introduction

Bovine leukemia virus (BLV) is a deltaretrovirus from the Orthoretrovirinae subfamily of the Retroviridae family. An essential step in the BLV replication cycle is the integration of DNA copy of its RNA genome into the DNA of a host cell [ 1 ]. Once integrated, the proviral DNA is replicated along with the host’s DNA during cellular divisions, as for any cellular gene. The BLV is the etiologic agent of enzootic bovine leukosis (EBL). BLV causes a persistent infection in cattle, and in most cases this infection is asymptomatic [ 2 ]. In one-third of infected animals the infection progresses to a state of persistent lymphocytosis, and in 1 to 10% of infected cattle it develops into lymphosarcoma [ 2 ]. BLV induces high economic losses due to trade restrictions, replacement cost, reduced milk production, immunosuppression, and increased susceptibility to pneumonia, diarrhea, mastitis, and so on [ 3 , 4 , 5 , 6 ]. BLV is globally distributed with a high prevalence, except for Western Europe and Oceania, where the virus has been successfully eradicated through detection and elimination of BLV-infected animals [ 7 , 8 ]. The agar gel immunodiffusion and ELISA for the detection of BLV-specific antibodies in sera and milk are the World Organization for Animal Health (WOAH, founded as OIE) prescribed tests for serological diagnosis but ELISA, due to its high sensitivity and ability to test many samples at a very low cost, is highly recommended [ 9 ]. Despite the advantages of serologic testing, there are some scenarios in which direct detection of the BLV genomic fragment was important to improve BLV detection. The most frequent cases is the screening of calves with maternal antibodies, acute infection, animals without persistent antibody response and animal subproducts (i.e., semen). In this regard, nucleic acid amplification tests such as real-time quantitative PCR (qPCR) allows for a rapid and highly sensitive detection of BLV proviral DNA (BLV DNA) that can be used to test infected and asymptomatic animals, before the elicitation of anti-BLV specific antibodies and when proviral load (PVL) are still low [ 10 ]. Furthermore, qPCR assays can serve as confirmatory tests for the clarification of inconclusive and discordant serological test results usually associated with these cases [ 11 ]. For these reasons, the inclusion of qPCR in combination with other screening tests might increase control programs efficiency. Additionally, qPCR allows the estimation of BLV PVL which is important for studying the dynamics of BLV infection (i.e., basic research). Further, considering that BLV PVL correlates with the risk of BLV transmission, this feature of qPCR can be exploited for developing rational segregation programs [ 12 , 13 ]. The results of Kobayashi et al. suggest that high PVL is also a significant risk factor for progression to EBL and should therefore be used as a parameter to identify cattle for culling from the herd well before EBL progression [ 14 ]. Several qPCRs have been developed globally for the quantitation of BLV DNA. Although most assays have been properly validated by each developer, a proper standardization and harmonization of such tests is currently lacking. Considering that standardization and harmonization of qPCR methods and results are essential for comparisons of data from BLV laboratories around the world, this could directly impact international surveillance programs and collaborative research. We built a global collaborative network of BLV reference laboratories to evaluate the interlaboratory variability of different qPCRs and sponsored a harmonization of assays to hopefully impact international surveillance programs and research going forward.

In 2018 we conducted the first global trial of this kind to assess the interlaboratory variability of six qPCRs for the detection of BLV DNA [ 15 ]. Since this complex process is a continuous rather than a one-time effort, we now started a second study of this type. In this follow up study, we built a more comprehensive sample panel, accounting for a broader geographical diversification. Additionally, we increased the number of participants to ten collaborating laboratories plus one WOAH reference lab and tested novel methodologies including digital PCR (ddPCR) and FRET-qPCR. Finally, we established the next steps towards the international standardization of molecular assays for the detection of BLV DNA.

Materials and methods

Participants.

The eleven laboratories that took part in the study were:(i) the Auburn University College of Veterinary Medicine (Auburn, Alabama, United States): (ii) AntelBio, a division of CentralStar Cooperative (Michigan, United States); (iii) Laboratórios Federais de Defesa Agropecuária de Minas Gerais (LFDA-MG, Pedro Leopoldo, Brasil); (iv) Centro de Investigación Veterinaria de Tandil (CIVETAN, Buenos Aires, Argentina); (v) the Faculty of Agriculture Iwate University (Iwate, Japan); (vi) Universidad de la República de Uruguay (UdelaR, Montevideo, Uruguay); (vii) the Croatian Veterinary Institute (Zagreb, Croatia); (viii) Instituto Nacional de Tecnología Agropecuaria (INTA, Buenos Aires, Argentina); (ix) Laboratorio Central de Veterinaria (LCV, Madrid, Spain); (x) the National Veterinary Research Institute (NVRI, Puławy, Poland) and (xi) the French Agency for Food, Environmental and Occupational Health and Safety (Anses, Niort, France). All European laboratories participating in this study are acting as national reference laboratories for EBL, NVRI acts as WOAH reference laboratory for EBL, while the remaining laboratories are nationally renowned entities for BLV diagnostics. The eleven participating methods are referred to below as qPCR1 – qPCR5, ddPCR6, qPCR7 – qPCR11, respectively.

Sample collection and DNA extraction

A total of 42 DNA samples obtained from blood of naturally BLV-infected dairy cattle from Poland, Moldova, Pakistan, Ukraine, Canada and United States were used for this study. Thirty-six of them were archival DNA samples obtained between 2012–2018 as described in our previous studies on samples from Poland ( n  = 21) [ 16 , 17 ], Moldova ( n  = 4) [ 18 ], Pakistan ( n  = 5) [ 19 ] and Ukraine ( n  = 6) [ 15 , 20 ]. Between 2020–2021 6 peripheral blood and serum samples from naturally BLV-infected cattle were obtained from three dairy farms of Alberta, Canada and two dairy farms of Michigan, US. Serological testing and sample processing were conducted by the laboratories from which the samples originated. The genomic DNA from Canadian and US samples was extracted from whole blood using a Quick DNA Miniprep Plus kit (Zymo Research) and a DNeasy Blood & Tissue Kit (Qiagen), respectively in University of Calgary and Michigan State University and sent to the NVRI in the form of DNA solutions. Additionally, one plasmid DNA sample (pBLV344) was kindly supplied by Luc Willems (University of Liège, Belgium) and DNA extracted from FLK-BLV cells were included as positive controls. Finally, DNA extracted from PBL of a serologically negative cattle was included as negative control. At the NVRI, the DNA concentration in all samples was estimated by spectrophotometry using a NanoPhotometer (Implen). Each sample was divided into eleven identical aliquots containing between 800 and 4,000 ng of lyophilised genomic DNA. Eleven identical sets of these samples were lyophilized (Alpha 1–4 LSC basic, Martin Christ Gefriertrocknungsanlagen GmbH) and distributed to participating laboratories. At the NVRI, all samples were coded (identification [ 21 ] run numbers 1 to 44) to perform a blinded testing. The samples, together with instructions for their preparation (Additional file 1), were shipped by air at room temperature (RT).

Examination of DNA quality/stability

Since different extraction methods and lyophilization process were employed for the preparation of the DNA samples, it was necessary to test the quality of the DNA at the NVRI laboratory. For that purpose, one complete set of samples ( n  = 44) was tested by Fragment Analyzer (Agilent Technologies), before and after freeze-drying, to assess DNA quality by calculating a Genomic Quality Number (GQN) for every sample. Low GQN value (< 2.5) represents sheared or degraded DNA. A high GQN (> 9) represents undegraded DNA. In addition, quality of DNA was assessed by determination of copy number of the histone H3 family 3A ( H3F3A ) housekeeping gene using quantitative real-time PCR (qPCR) [ 22 ]. The qPCR results were expressed as the number of H3F3A gene copies per 300 ng of DNA in each sample. Grubbs´ test was performed to determine outliers. To test the stability of DNA, samples were stored for 20 days at RT (10 days) and at + 4 °C (10 days) and were retested by Fragment Analyzer and qPCR 21 days later. A Mann–Whitney U-test was used to compare the median values between fresh and stored samples (time 0 and time 1), respectively.

Description of BLV qPCR protocols used by participating laboratories

All participating laboratories performed their qPCR or ddPCR using a variety of different equipment, reagents, and reaction conditions, which had been set up, validated, and evaluated previously and are currently used as working protocols. The specific features of each of these protocols are described below and summarized in Table  1 .

All laboratories applied standard procedures for avoiding false-positive results indicative of DNA contamination, such as the use of separate rooms for preparing reaction mixtures, adding the samples, and performing the amplification reaction. One of the ten BLV qPCRs used LTR region and the remaining nine qPCRs used the pol gene as the target sequence for amplification, while the ddPCR amplified the env gene.

Method qPCR1

The BLV qPCR amplifying a 187-bp pol gene was performed according to a previously published methods [ 23 , 24 ]. A real-time fluorescence resonance energy transfer (FRET) PCR was carried out in a 20-μl PCR mixture containing 10 μl handmade reaction master mix and 10 μl genomic DNA. The PCR buffer was 4.5 mM MgCl2, 50 mM KCl, 20 mM Tris–HCl, pH 8.4, supplemented with 0.05% each Tween20 and Non-idet P-40, and 0.03% acetylated BSA (Roche Applied Science). For each 20 μl total reaction volume, the nucleotides were used at 0.2 mM each and 1.5 U Platinum Taq DNA polymerase (Invitrogen, Carlsbad, CA, USA) was used. Primers were used at 1 μM, LCRed640 probe was used at 0.2 μM, and 6-FAM probe was used at 0.1 μM. Amplification was performed in the Roche Light Cycler 480 II (Roche Molecular Biochemicals) using 10 min denaturation step at 95 °C, followed by 18 high-stringency step-down thermal cycles and 30 low-stringency fluorescence acquisition cycles.

A plasmid containing the BLV-PCR amplicon region was diluted ten-fold from 1 × 10 5 copies to 10 copies per 10 µl and was used as a standard to measure the BLV copy numbers.

Method qPCR2

A BLV proviral load qPCR assay developed by AntelBio, a division of CentralStar Cooperative Inc. on Applied Biosystems 7500 Real-Time PCR system [ 25 , 33 ]. This multiplex assay amplifies the BLV pol gene along with the bovine β-actin gene and an internal amplification control, “Spike”. A quantitative TaqMan PCR was carried out in a 25-μl PCR mixture containing 12.5 µl of 2X InhibiTaq Multiplex HotStart qPCR MasterMix (Empirical Bioscience), 16 nM each BLV primer, 16 nM each β-actin primer, 8 nM each spike primer, 8 nM BLV FAM-probe, 8 nM β-actin Cy5-probe, 4 nM spike JOE-probe, 1 µl of an internal spike-in control (10,000 copies per µl), 7.25 µl of nuclease-free water and 4 µl of DNA sample for each qPCR reaction. The thermal PCR protocol was as follows: 95 °C for 10 min, 40 × (95 °C for 15 s, 60 °C for 1 min). Copy numbers of both the BLV pol gene and bovine β-Actin were derived using a plasmid containing target sequences, quantified by ddPCR, diluted 1 × 10 6 copies per µl to 10 copies per µl in tenfold dilutions. DNA concentrations of each sample were measured using a Qubit 4 Fluorometer and used in combination with the qPCR copy numbers to calculate BLV copies per 100 ng.

Method qPCR3

The qPCR assays for the BLV LTR gene were performed according to a previously published methods [ 26 ]. Genomic DNA was amplified by TaqMan PCR with 10 μl of GoTaq Probe qPCR Master Mix × 2 (Promega), 0.6 pmol/μl each primer, 0.3 pmol/µl double-quenched probe and 100 ng genomic DNA. Amplification was performed in the CFX96 cycler (BioRad) according to the protocol: 5 min denaturation at 95°C followed by 45 cycles (60 s at 94°C and 60 s at 60°C). The efficiency of each reaction was calculated from the serial dilution of DNA extracted from BLV persistently infected fetal lamb kidney (FLK) cells, starting at a concentration of 100 ng/µl [ 21 ]. The detection limit was tested using a plasmid containing the target of the qPCRs, starting at 10 3 ng/µl.

Method qPCR4

The quantitative real-time PCR was done with the primers for the BLV pol gene as previously described [ 34 ]. The qPCR reaction mix contained 1 × PCR Master Mix with SYBR Green (FastStart Universal SYBR Green Master Rox, Roche), 0.3 μM each primer and 30 ng of extracted genomic DNA. Amplification was performed in QuantStudio 5 Real-Time PCR System (Applied Biosystems) under the following conditions: 2 min at 50 °C, 10 min at 95 °C, 40 cycles of 15 s at 95 °C and 60 s at 60 °C. A standard curve of six tenfold serial dilutions of pBLV, containing 1 × 10 6 to 10 BLV copies, was built and run 3 times for validation of the method. The number of provirus copies per reaction (100 ng) was calculated.

Method qPCR5

BLV PVLs were determined by using qPCR kit, RC202 (Takara Bio, Shiga, Japan) [ 28 , 35 ]. This qPCR assay amplifies the BLV pol gene along with the bovine RPPH1 gene as an internal control. Briefly, 100 ng genomic DNA was amplified by TaqMan PCR with four primers for pol gene and RPPH1 gene according to the manufacturer’s instructions: 30 s denaturation at 95 °C followed by 45 cycles (5 s at 95 °C and 30 s at 60 °C). The qPCR was performed on a QuantStudio 3 Real-Time PCR System (Thermo Fisher Scientific K.K., Tokyo, Japan). Standard curve was generated by creating tenfold serial dilutions of the standard plasmid included in the kit. The standards for calibration ranged from 1 to 10 6 copies/reaction and were run in duplicate. The number of provirus copies per 100 ng was calculated.

Method ddPCR6

The digital droplet PCR (ddPCR) assay for the env gene of the BLV was performed using the protocol previously described by [ 28 , 29 ]. An absolute quantification by TaqMan ddPCR was performed in a typical 20-μl assay, 1 μl of DNA sample was mixed with 1 μl of each primer (10 μM), 0.5 μl of probe (10 μM), and 2 × Supermix emulsified with oil (Bio-Rad). The droplets were transferred to a 96-well plate (Eppendorf). The PCR assay was performed in a thermocycler (C1000 touch cycler; Bio-Rad) with the following parameters: initial denaturation of 10 min at 95 °C, then 40 cycles of 30 s at 94 °C, and 1 min at 58 °C, with final deactivation of the enzyme for 10 min at 98 °C. The presence of fluorescent droplets determined the number of resulting positive events that were analyzed in the software (QuantaSoft v.1.7.4; Bio-Rad), using dot charts. The number of provirus copies per 100 ng were calculated. Each sample was run in duplicate, and results were averaged.

Method qPCR7

This qPCR method for the BLV pol gene is a modified option of widely available quantitative TaqMan qPCR described by Rola-Łuszczak et al. [ 11 ], using the same primers and standards. A quantitative TaqMan PCR was performed in a 20 μl PCR mix containing 10 μl of 2 × ORA qPCR Probe ROX L Mix (highQu, Kraichtal, Germany), 2 μl primer/probe mix (final concentration 400 nM of each of the primers, 200 nM of BLV probe), and 3 μl extracted genomic DNA. Amplification was performed in the Rotor-Gene Q system (Qiagen) with an initial denaturation step and polymerase activation at 95 °C for 3 min, followed by 45 cycles of 95 °C for 5 s and 60 °C for 30 s. As a standard, plasmid pBLV1 (NVRI, Pulawy, PL) containing a BLV pol fragment was used. Tenfold dilutions of plasmid DNA were made from 1 × 10 10 copies to 1 × 10 1 copies per reaction and used to generate the standard curve and estimate BLV copy number per 100 ng.

Method qPCR8

Proviral load quantification was assessed by SYBR Green real-time quantitative PCR (qPCR) using the pol gene as the target sequence [ 36 ]. Briefly, 12-μl PCR mixture contained Fast Start Universal SYBR Green Master Mix (Roche), 800 nM each BLV pol primers and 1 µl DNA as template. The reactions were incubated at 50 °C for 2 min and 95 °C for 10 min, followed by 40 cycles at 95 °C for 15 s, 55 °C for 15 s and 60 °C for 1 min. All samples were tested in duplicate on a StepOne Plus machine (Applied Biosystems). A positive and negative control, as well as a no-template control, were included in each plate. After the reaction was completed, the specificity of the amplicons was checked by analyzing the individual dissociation curves. As a standard, plasmid pBLV1 (NVRI, Pulawy, PL) containing a BLV pol fragment was used. Tenfold dilutions of plasmid DNA were made from 1 × 10 6 to 10 copies per µl and used to generate the standard curve and estimate BLV copy number per 100 ng.

Method qPCR9

This qPCR method is a modified option of widely available quantitative TaqMan qPCR described by Rola-Łuszczak et al. [ 11 ], using the same primers and standards. The detection of BLV genome was combined with an endogenous control system (Toussaint 2007) in a duplex assay. Briefly, 20-µl qPCR reaction contained AhPath ID™ One-Step RT-PCR Reagents with ROX (Applied Biosystems, CA, USA) – 10 µl of 2 × RT-PCR buffer and 0.8 µl of 25 × RT-PCR enzyme mix, 400 nM each primer for pol gene, 100 nM BLV specific probe, 40 nM each β-actin primer, 40 nM β-actin specific probe and 2 µl DNA sample. All samples were tested in ABI7500 Real-Time PCR System (Applied Biosystems) according to the following protocol: 10 min at 48 °C (reverse transcription), 10 min at 95 °C (inactivation reverse transcriptase / activation Taq polymerase) followed by 45 cycles (15 s at 95 °C and 60 s at 60 °C). As a standard, plasmid pBLV1 (NVRI, Pulawy, PL) containing a BLV pol fragment was used. Tenfold dilutions of plasmid DNA were made from 1 × 10 4 copies to 0.1 copies per μl and used to generate the standard curve and estimate BLV copy number per 100 ng.

Method qPCR10

The BLV qPCR was performed as published previously [ 11 ]. A quantitative TaqMan PCR was carried out in a 25-μl PCR mixture containing 12.5 μl of 2 × QuantiTect Multiplex PCR NoROX master mix (Qiagen), 0.4 μM each primer, 0.2 μM specific BLV probe, and 500 ng of extracted genomic DNA. Amplification was performed in the Rotor-Gene Q system (Qiagen) using an initial denaturation step and polymerase activation at 95 °C for 15 min, followed by 50 cycles of 94 °C for 60 s and 60 °C for 60 s. All samples were amplified in duplicate. As a standard, the pBLV1 plasmid (NVRI, Pulawy, PL), containing a 120-bp BLV pol fragment, was used. Tenfold dilutions of this standard were made from 1 × 10 6 copies per μl to 100 copies per μl and were used to estimate the BLV copy numbers per 100 ng.

Method qPCR11

This qPCR method for the BLV pol gene is a modified option of widely available quantitative TaqMan qPCR described by Rola-Łuszczak et al. [ 11 ], using the same primers and standards. The reaction mixture contained 400 nM of each primer, 200 nM of probe, 10 µl of 2 × SsoFast probes supermix (Bio-Rad), 5 µl of DNA sample and H 2 O up to 20 µl of the final volume. PCR assays were carried out on a CFX96 thermocycler (Bio-Rad) under the following amplification profile: 98 °C for 3 min, followed by 45 cycles of 95 °C for 5 s and 60 °C for 30 s. As a standard, plasmid pBLV1 (NVRI, Pulawy, PL) containing a BLV pol fragment was used. Tenfold dilutions of plasmid DNA were used to generate the standard curve and estimate BLV copy number per 100 ng.

Analysis of BLV pol, env and LTR sequences targeted by particular qPCR/ddPCR assays

In order to assess full-length pol , env and LTR sequence variability among BLV genotypes, all BLV sequences ( n  = 2191) available on 30 September 2023 in GenBank ( https://www.ncbi.nlm.nih.gov/GenBank/ ) repository were retrieved. From the collected sequences, 100 pol , env and LTR sequences, which were characterized by the highest level of sequence variability and divergence, were selected for the further analysis. A pol -based, env -based and LTR-based maximum likelihood (ML) phylogenetic trees (see Additional file 6) was constructed to assign genotypes to the unassigned BLV genomes [ 37 , 38 , 39 ]. For all genes and LTR region the Tamura-Nei model and Bootstrap replications (1,000) were applied. In this analysis, pol sequences were assigned to 7 BLV genotypes (G1, G2, G3, G4, G6, G9, and G10), while env and LTR sequences were assigned to 10 BLV genotypes (G1, G2, G3, G4, G5, G6, G7, G8, G9, and G10). Phylogeny of the same isolates assigned to particular genotypes by ML method was confirmed by Mr. Bayes analysis [ 40 , 41 , 42 ] (data not shown). From this analysis, a total of 100 full-length pol, env and LTR sequences were used for multiple-sequence alignment (MSA) using ClustalW algorithm, implemented in MEGA X. For all sequences, nucleotide diversity (π), defined as the average number of nucleotide differences per site between two DNA sequences in all possible pairs in the sample population, was estimated using MEGA X. To measure the relative variation in different positions of aligned genes and LTR region the Shannon’s entropy (a quantitative measure of diversity in the alignment, where H = 0 indicates complete conservation) was estimated using BioEdit v. 7.2.5 software 64. The statistical analyses were performed using DATAtab e.U. Graz, Austria and GraphPad Software by Dotmatics, Boston.

Examination of the quality and stability of DNA samples

To test the quality of DNA samples, the H3F3A copy number of each individual sample was assessed by qPCR at the NVRI. Copy numbers were normalized to DNA mass input and results were expressed as copy numbers per 300 ng of total DNA. The respective values were tested by Grubbs' test. The results for 43 DNA samples (sample ID: 42 with BLV genome plasmid was excluded) followed a normal distribution (Shapiro–Wilk 0.97; P  = 0.286), with a mean value of 35,626 copies (95% confidence interval [ 43 ] 33,843 to 37,408 copies), a minimum value of 19,848 copies and a maximum value of 46,951 copies (see Additional file 2). Despite a low value for sample ID: 40 no significant outlier was detected in the dataset ( P  > 0.05). Therefore, it can be assumed that the DNA quality was acceptable for all samples present in the panel. Next, DNA stability was assessed by retesting the H3F3A copy numbers in each sample ( n  = 43) after a combined storage consisting in 10 days at RT and 10 days at + 4°C. A Mann–Whitney U-test was used to compare the median values between fresh and stored samples (time 0 and time 1, respectively), and no significant difference was observed at the 5% level ( P  = 0.187) (Fig.  1 A).

Assessment of the stability of DNA samples. A Shown are copy numbers of the H3F3A housekeeping gene in 43 DNA samples that were stored in 10 days at RT and 10 days at + 4°C and tested twice with a 21-day interval. A Mann–Whitney U-test was used to compare the median values between two groups ( P  = 0.187); B Shown are GQN values ( n  = 43) tested twice with a 21-day interval: `before freeze-drying` and `after freeze-drying`. A Mann–Whitney U-test results between two groups ( P  = 0.236)

In addition, the quality of DNA samples after lyophilization was analyzed. DNA from individual samples ( n  = 43) was assessed with the genomic DNA quality number on the Fragment Analyzer system. The GQN from all lyophilized samples ranged from 4.0 to 9.7—that represented undegraded DNA. There was no significant difference in GQN values between `before freeze-drying` and `after freeze-drying` groups with respect to the corresponding DNA samples ( P  = 0.236) (Fig.  1 B). Altogether, these results suggested that sample storage, lyophilization and shipping has a minimal impact in DNA stability and further testing during the interlaboratory trial.

Detection of BLV proviral DNA by different qPCR assays

A total of 44 DNA samples, including two positive (ID: 42 and 43) and one negative (ID: 32) controls, were blinded and independently tested by eleven laboratories using their own qPCR methods (Table  2 ). All laboratories measured the concentration of DNA in samples (Additional file 3). BLV provirus copy number was normalized to DNA concentration and expressed per 100 ng of genomic DNA for each test.

Except for the positive (pBLV344 and FLK cell line) and the negative controls, all samples had previously shown detectable levels of BLV-specific antibodies (BLV-Abs) by enzyme-linked immunosorbent assays (ELISA). During the current interlaboratory study, both the positive and negative controls were assessed adequately by all eleven PCR tests. Of all 43 positive samples, 43, 35, 37, 36, 40, 32, 40, 42, 42, 42 and 41 samples were detected as positive by the qPCR1, qPCR2, qPCR3, qPCR4, qPCR5, ddPCR6, qPCR7, qPCR8, qPCR9, qPCR10 and qPCR11 methods, respectively. Based on these observations, the most sensitive method was the qPCR1, and the method with the lowest sensitivity was the ddPCR6. Twenty-nine out of 44 samples were identified correctly by all qPCRs. The remaining 15 samples gave discordant results. Comparison of qualitative results (positive versus negative) from all eleven methods revealed 87.33% overall agreement and a kappa value of 0.396 (Cohen's kappa method adapted by Fleiss) [ 44 , 45 ]. The levels of agreement among the results from the eleven methods are represented in Table  3 . The maximum agreement was seen between two methods (qPCR9 and qPCR10 [100% agreement and a Cohen's kappa value of 1.000]) that used similar protocols and targeted the same region of BLV pol .

Analysis of BLV pol, env and LTR sequences targeted by particular PCR assays

Due to differences in performance observed among the pol -based qPCR assays (the qPCR1, qPCR2, qPCR4, qPCR5 and qPCR7- qPCR11 methods), and considering that the env -based ddPCR6 and LTR-based qPCR3 assay showed the lowest sensitivity and the poorest agreement with the other assays, the degree of sequence variability between the pol , env and LTR genes was addressed. From the MSAs for pol , env and LTR, the nucleotide diversity (π) was calculated. The π value for pol gene was lower than that for LTR and env gene (π pol , 0.023 [standard deviation {SD}, 0.018]; π LTR , 0.024 [SD, 0.011]; π env , 0.037 [SD, 0.013]). From this analysis, pol sequences appeared to be less variable than env and LTR sequences. In addition, we performed a Shannon entropy-based per-site variability profile of the pol , env and LTR sequences used in this study (Fig.  2 A-C).

Sequence variability measured as per-site entropy. A Multiple alignment of the pol gene showing the locations of qPCR fragments in regions of the pol gene for the qPCR1 (highlighted in pink), qPCR4 (highlighted in yellow) and for the qPCR7, qPCR8, qPCR9, qPCR10 and qPCR11 assays (highlighted in orange). B Multiple alignment of the env gene targeted by ddPCR6 (highlighted by blue rectangle). C Multiple alignment of the LTR region by qPCR3 (highlighted in mint)

The all-observed entropy plots were homogeneous along the whole sequences. Considering the three regions of pol gene, the highest entropy (4.67) occurred in the region targeted by the qPCR1 primers, whereas the entropy for qPCR7—qPCR11 and qPCR4 primers were 1.57 and 0.38, respectively. For the LTR region targeted by qPCR3 primers and for env gene targeted by ddPCR6, the total entropy was equal to 4.46 and 7.85, respectively. This analysis showed a marked region of variability for LTR and env fragments. Interestingly, we noted that the qPCR7—qPCR11 targeted the most conserved regions of reverse transcriptase and qPCR4 primers targeted the most-conserved region of virus integrase (Fig.  2 A-C; see also Additional file 7).

Quantitation of BLV proviral DNA by different qPCR/ddPCR assays

To analyze whether the range of copy numbers detected by each qPCR was comparable to those of the others, Kruskal–Wallis one-way analysis of variance (ANOVA) was used. The violin plots were used to visualize the ANOVA results (Fig.  3 A-B).

Comparison of detection of BLV proviral DNA copy numbers by eleven testing methods. Shown is a box plot of data from Kruskal–Wallis ANOVA, a rank test. The DNA copy numbers for 41 samples, determined independently by each of the 11 qPCRs, were used for the variance analysis. In this analysis, the positive controls (sample ID 42 and ID 43) and negative control (sample ID 32) were excluded. A Violin plot for graphical presentation of the ANOVA of proviral copy number values. B Violin plot for ANOVA analysis of variance, copy number values are presented on a logarithmic scale (Log1.2) for better illustration of copy number differences between PCR methods

The grouping variable revealed significant differences among the distributions of proviral DNA copy numbers with the various qPCRs ( P  < 0.001). These results showed that the abilities of qPCRs/ddPCR to determine the proviral DNA copy number differed. A Dunn-Bonferroni test was used to compare the groups in pairs to find out which was significantly different. The Dunn-Bonferroni test revealed that the pairwise group comparisons of qPCR2—qPCR4, qPCR3—ddPCR6, qPCR4—qPCR5, qPCR4—ddPCR6, qPCR4—qPCR9, qPCR4—qPCR10, qPCR5—qPCR11, ddPCR6—qPCR11 and qPCR9—qPCR11 have an adjusted P value less than 0.05 and thus, it can be assumed that these groups were significantly different in each pair (see Additional file 4). The Pareto chart was used to show the average copy number values of all methods in descending order. These Pareto charts were prepared based on 80–20 rule, which states that 80% of effects come from 20% of the various causes [ 46 ]. The methods that generated the highest copy numbers was qPCR3 and qPCR4, on the other hand the lowest copy numbers and/or highest negative results were generated by ddPCR6 (Fig.  4 ).

A Pareto chart with the proviral BLV copy mean values for eleven PCR assay arranged in descending order. Pareto charts was prepared based on 80–20 rule, which states that 80% of effects come from 20% of the various causes

The correlations between copy numbers detected by different qPCRs and ddPCR assays were calculated. The Kendall's Tau correlation coefficient measured between each pair of the assays was shown in the Additional file 5 and in Fig.  5 as a correlation heatmap. The average correlation for all qPCRs and ddPCR assays was strong (Kendall's tau = 0.748; P  < 0.001).

The heatmap of Kendall’s tau correlation coefficients between copy numbers detected by ten qPCRs and one ddPCR. Statistically significant differences in the distribution of copy numbers, a moderate, strong and very strong correlation between particular qPCRs/ddPCR was observed. The strength of the association, for absolute values of r, 0–0.19 is regarded as very weak, 0.2–0.39 as weak, 0.40–0.59 as moderate, 0.6–0.79 as strong and 0.8–1 as very strong correlation

Since the differences between PCR tests may be influenced by the number of BLV proviral copies present in each sample, we compared the average number of BLV copies between a group of genomic DNA samples that gave concordant results (group I [ n  = 28]) and a group that gave discordant results (group II [ n  = 15]). The mean number of copies was 73,907 (minimum, 0; maximum, 4,286,730) in group I, and 3,479 (minimum, 0; maximum, 218,583) in group II, and this difference was statistically significant ( P  < 0.001 by a Mann–Whitney U- test) (Fig.  6 ).

Impact of BLV proviral copy numbers on the level of agreement. Violin plot for graphical presentation of Mann–Whitney U test. The test was performed to compare BLV provirus copy number in two groups of samples: 28 samples with fully concordant results from all eleven qPCR/ddPCR assays (left) and 15 samples with discordant results from different qPCR/ddPCR assays (right) ( P  < 0.001). Sample ID 42 was excluded from the statistical analysis

The results show that the concordant results group had considerably higher copy numbers (median, 5,549.0) than the discordant results group (median, 6.3).

BLV control and eradication programs consist of correct identification and subsequent segregation/elimination of BLV-infected animals [ 47 ]. Detection of BLV- infected cows by testing for BLV-specific antibodies in serum by agar gel immunodiffusion and ELISA is the key step and standard to be implemented of EBL eradication programs according to WOAH ( https://www.woah.org/en/disease/enzootic-bovine-leukosis/) [ 9 ]. Despite the low cost and high throughput of serological tests, there are several scenarios where highly specific and sensitive molecular assays for the detection of BLV DNA might improve detection and program efficiency.

In this perspective, qPCR assays can detect small quantities of proviral DNA during acute infection, in which animals show very low levels of anti-BLV antibodies [ 43 , 48 , 49 , 50 ]. qPCR methods can also work as confirmatory tests to clarify ambiguous and inconsistent serological test results [ 11 ]. Such quantitative features of qPCRs are crucial when eradication programs progress and prevalence decreases. Moreover, qPCR allows not only the detection of BLV infection but also estimation of the BLV PVL, which directly correlates with the risk of disease transmission [ 51 , 52 ]. This feature of qPCR allows for a rational segregation of animals based on the stratified risk of transmission. These considerations allow for greater precision in the management of BLV within large herds with a high prevalence of BLV ELISA-positive animals to effectively reduce herd prevalence [ 13 , 53 ]. BLV is a global burden and the lack of technical standardization of molecular detection systems remains a huge obstacle to compare surveillance data globally based on the first interlaboratory trial performed in 2018 [ 15 ]. In the 2018 study we observed an adjusted level of agreement of 70% comparing qualitative qPCR results; however, inconsistencies amongst methods were larger when low number of copies of BLV DNA were compared. Samples with low copies of BLV DNA (< 20 copies per 100 ng) accounted for the higher variability and discrepancies amongst tests. We concluded from the first interlaboratory trial that standardizing protocols to improve sensitivity of assays with lower detection rates was necessary.

In this follow up study, we re-tested the TaqMan BLV qPCR developed and validated by NVRI (acting as reference WOAH laboratory) and the one adapted from this original protocol to be used with SYBR Green dye, allowing a significant reduction in costs [ 11 ]. Another 3 laboratories also performed NVRI´s qPCR with slight modifications (i.e., Spain performed a multiplex assay for internal normalization). The remaining 6 labs introduced novel methodologies to the trial including one ddPCR (UY).

To compare different qPCR methods, a more comprehensive sample panel, accounting for a more geographical diversification was used in this trial. The amounts of BLV DNA in these samples were representative of the different BLV proviral loads found in field samples (from 1 to > 10,000 copies of BLV proviral DNA). Of note, 34% of reference samples had less than 100 copies of BLV DNA per 100 ng; samples were lyophilized to grant better preservation and reduced variability during distribution to participants around the globe.

The panel included a single negative control and two positive controls. Diagnostic sensitivity (DxSn) was estimated for each qPCR. Considering the 43 positive samples, the DxSn for the different qPCRs were: qPCR1 = 100%, qPCR2 = 82%, qPCR3 = 86%, qPCR4 = 84%, qPCR5 = 93%, ddPCR6 = 74%, qPCR7 = 93%, qPCR8 = 98%, qPCR9 = 98%, qPCR10 = 98% and qPCR11 = 95%. The most sensitive method was the qPCR1, and the method with the lowest sensitivity was the ddPCR6 method. Twenty-nine out of 44 samples were identified correctly by all qPCRs. The remaining 15 samples gave discordant results. The comparison of qualitative qPCR results among all raters revealed an overall observed agreement of 87%, indicating strong interrater reliability (Cohen´s kappa = 0.396) [ 54 , 55 ].

There are several factors that contribute to variability in qPCR results (i.e., number of copies of target input, sample acquisition, processing, storage and shipping, DNA purification, target selection, assay design, calibrator, data analysis, etc.). For that reason and as expected, the level of agreement among sister qPCRs (qPCR7, qPCR9-11) sharing similar protocols was higher compared to the rest of assays; this was also true for qPCR8 which targets the same region of BLV pol gene (shares same primers) but has a particular set-up to be used with SYBR Green chemistry. Oppositely, lower sensitivity and larger discrepancy against other tests was observed for the ddPCR6 and qPCR2-4.

Based on these observations we investigated which factors might have accounted for larger assessment variability amongst tests. In the first place, we observed that the use of different chemistries was not detrimental for the sensitivity and agreement among tests; similar DxSn and comparable level of agreement were obtained comparing TaqMan (qPCR7, 10, 11) vs SYBR Green (qPCR8) chemistries while targeting identical BLV sequence and using same standards. Also, when a multiplex qPCR (TaqMan) targeting the same BLV sequence and using the same standard was compared to previous ones, agreement was kept high, indicating that the lower sensitivity described for some multiplex qPCRs did not take place in this comparison. The use of an international calibrator and the efficiency estimation (standard curve) might inform variability associated with different chemistries. In contrast, another multiplex assay targeting another region of BLV pol (qPCR2) showed much lower sensitivity and agreement. As qPCR2 is performed as service by private company and oligonucleotide sequences were not available, we were not able to investigate in which proportion each of these two variables contributed to the lower performance of this assay, but we note the addition of 4 µl genomic DNA to this assay that would have an impact the DxSn. In this regard, there is substantial evidence showing that the variability of target sequence among strains from different geographical areas, might affect the sensitivity of BLV qPCRs. Previous studies comparing the pol , gag , tax and env genes reported that the pol gene was the most suitable region to target for diagnostic purposes, since it provided the most-sensitive assays [ 11 , 15 , 56 , 57 , 58 , 59 ]. This might be due in part to higher sequence conservation of pol among strains from different geographical areas. Supporting this observation, it is noticeable how JPN qPCR improved their performance in the current trial, by targeting pol in place of tax , as it did in the previous interlaboratory trial. Since it is a commercial test, we cannot exclude other factors contributing for the performance upgrade observed for this qPCR. In the current study, qPCR3 and ddPCR6 targeting LTR and env sequences, showed lower performances than other assays. Standardization of DNA input into each qPCR would have likely resulted in higher concordance in results. For instance, qPCR1 added 10 µl of genomic DNA per reaction and ddPCR6 added 1 µl of genomic DNA, impacting the resulting sensitivity differences.

Since the sensitivity of each assay and, consequently, the level of agreement among assays might also be influenced by the number of BLV DNA copies present in each sample [ 48 ], we compared the average number of BLV DNA copies between a group of genomic DNA samples that gave concordant results and a group that gave discordant results, and observed that samples that gave discordant results had significantly lower numbers of BLV DNA copies than samples that gave concordant results. Related to this point, the degradation of target DNA during lyophilization, shipment and resuspension, could have been more significant in low-copy compared to high-copy samples. Consequently, the degradation of target DNA in samples with low copies of BLV DNA might have accounted for the greater level of discrepancy within this subset of samples. The rational of adding a large proportion of such samples (34% samples with less than 100 BLV copies per 100 ng of total DNA) was to mimic what is frequently observed in surveillance programs (i.e., hyperacute infection, chronic asymptomatic infection, etc.).

Quantitative methods for the detection of BLV DNA copies are important for segregation programs based on animal level of BLV PVL, as well as for scientific research and the study of BLV dynamics. When the numbers of copies of BLV DNA detected by different assays were compared, in the present study, we observed that although the ability to quantify BLV DNA differed among qPCRs/ddPCR and there were statistically significant differences in the distribution of copy numbers among assays, a strong average correlation was found for the eleven qPCRs/ddPCR. In this regard, the lack of an international calibrator (standard curve) could be a major contributor to the increment of quantitative variation amongst laboratories. For that reason, plasmid pBLV1 containing pol 120 bp sequence was originally constructed for use as standard for quantification and shared with some collaborators (i.e., qPCR7, qPCR8, qPCR 9, qPCR10 and qPCR11). Remarkably, the laboratories used pBLV1 standard in the current trial obtained the most comparable results, indicating that the use of an international standard may have significant impact on the convergence of results; such standard reference material should be prepared under identical conditions. To avoid further variability a detailed protocol for lyophilized DNA sample resuspension, quantitation and template input into each qPCR should be shared with all participants.

Conclusions

BLV DNA was detected with different level of sensitivity in serologically positive samples from different origin and classified into different BLV genotypes. Overall agreement was high; however, we found significant differences in results for the samples with low BLV DNA copy numbers. This second interlaboratory study demonstrated that differences in target sequence, DNA input and calibration curve standards can increase interlaboratory variability considerably. Next steps should focus on (i) standard unification (international gold standard) to estimate individual test efficiency and improve quantitative accuracy amongst tests; (ii) building a new panel of samples with low BLV DNA copy numbers to re-evaluate sensitivity and quantitation of molecular methods. Since no variation was observed in samples from different genotypes, all samples will be collected in Poland to standardize the collection, purification, lyophilization and shipping steps with precise instructions for suspension and constant input volume for the PCR reaction. Finally, we believe that following this standardization approach we will be able to improve overall agreement amongst tests, improving the diagnostic of BLV around the world.

Availability of data and materials

Not applicable.

Data availability

No datasets were generated or analysed during the current study.

Abbreviations

One-way analysis of variance

Bovine leukemia virus

BLV-specific antibodies

Digital PCR

Diagnostic sensitivity

Enzootic bovine leukosis

Enzyme-linked immunosorbent assays

Real-time fluorescence resonance energy transfer PCR

Genomic quality number

Histone H3 family 3A housekeeping gene

Maximum likelihood phylogenetic tree

Multiple-sequence alignment

Peripheral blood leukocytes

Phosphate-buffered saline

Proviral load

Quantitative real-time PCR

Room temperature

World Organisation for Animal Health

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Acknowledgements

The authors thank Luc Willems (University of Liège, Belgium) for plasmid DNA sample pBLV344; Marlena Smagacz and Eliza Czarnecka (National Veterinary Research Institute, Poland) for lyophilizing DNA samples and DNA analysis, respectively; Ali Sakhawat (Animal Quarantine Department, Pakistan), Vitaliy Bolotin (National Scientific Center IECVM, Ukraine), Frank van der Meer and Sulav Shrestha (University of Calgary, Canada) for sharing material.

The APC was funded by the National Veterinary Research Institute, Puławy, Poland.

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Department of Biochemistry, National Veterinary Research Institute, Puławy, 24-100, Poland

Aneta Pluta & Jacek Kuźmak

Instituto de Virología E Innovaciones Tecnológicas (IVIT), Centro de Investigaciones en Ciencias Veterinarias y Agronómicas (CICVyA), Instituto Nacional de Tecnología Agropecuaria (INTA) - CONICET, Buenos Aires, Argentina

Juan Pablo Jaworski & Vanesa Ruiz

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Casey Droscha & Sophie VanderWeele

Department of Animal Science, College of Agriculture and Natural Resources, Michigan State University, East Lansing, Michigan, 48824, USA

Tasia M. Taxis

Niort Laboratory, Unit Pathology and Welfare of Ruminants, French Agency for Food, Environmental and Occupational Health and Safety (Anses), Ploufragan-Plouzané, Niort, France

Stephen Valas

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Contributions

Proposed the conception and design of the study, A.P.; data curation, A.P., J.P.J., C.D., S.V., D.B., A.S., K.M., R.P., G.D., M.F.C. and CH.W.; investigation, A.P., V.R., S.VW., S.V., A.J., M.J.R., K.N., M.L.B., M.L.G., P.L., A.F., A.G. and S.B., formal analysis, A.P.; statistical analysis, A.P.; database analysis, A.P., visualization of the results, A.P.; resources, A.P., T.M.T. and J.K; writing—original draft preparation, A.P., J.P.J.; writing—review and editing, A.P., J.P.J., C.D., S.VW., T.M.T. and J.K; project administration, A.P. All authors read and approved the submitted version.

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Correspondence to Aneta Pluta .

Ethics declarations

Ethics approval and consent to participate.

The study was approved by the Veterinary Sciences Animal Care Committee No. AC21-0210, Canada; the Institutional Animal Care and Use Committee No. PROTO202000096 from 4/13/2020 to 4/14/2023, Michigan State University, United States and the Ethics Review Board, COMSATS Institute of Information Technology, Islamabad, Pakistan, no. CIIT/Bio/ERB/17/26. Blood samples from Polish, Moldovan and Ukrainian cattle, naturally infected with BLV, were selected from collections at local diagnostic laboratories as part of the Enzootic bovine leukosis (EBL) monitoring program between 2012 and 2018 and sent to the National Veterinary Research Institute (NVRI) in Pulawy for confirmation study. The approval for collection of these samples from ethics committee was not required according to Polish regulation (“Act on the Protection of Animals Used for Scientific or Educational Purposes”, Journal of Laws of 2015). All methods were carried out in accordance with relevant guidelines and regulations. The owners of the cattle herds from which the DNA samples originated, the district veterinarians caring for these farms and the ministries of agriculture were informed and consented to the collection of blood from the animals for scientific purposes and the sending of samples to NVRI.

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Supplementary Information

12917_2024_4228_moesm1_esm.pdf.

Additional file 1. Copy of the instruction included with the panel of 44 DNA samples sent to participating laboratories for dilution of the lyophilisates

12917_2024_4228_MOESM2_ESM.png

Additional file 2. Detection of the H3F3A gene copy number in 43 DNA samples; no outlier was found for any samples ( P <0.05) (two-sided).

12917_2024_4228_MOESM3_ESM.docx

Additional file 3. Concentration values of 44 DNA samples measured by the 11 participating laboratories (given in ng per µl)

12917_2024_4228_MOESM4_ESM.pdf

Additional file 4. Post hoc - Dunn-Bonferroni-Tests. The Dunn-Bonferroni test revealed that the pairwise group comparisons of qPCR2 - qPCR4, qPCR3 - ddPCR6, qPCR4 - qPCR5, qPCR4 - ddPCR6, qPCR4 - qPCR9, qPCR4 - qPCR10, qPCR5 - qPCR11, ddPCR6 - qPCR11 and qPCR9 - qPCR11 have an adjusted p-value less than 0,05

12917_2024_4228_MOESM5_ESM.docx

Additional file 5. Kendall's Tau correlation coefficient values measured between each pair of assays. The numbers 1 to 11 in the first column and last row of the table indicate the names of the assays qPCR1-qPCR5, ddPCR6, qPCR7-qPCR11 respectively

12917_2024_4228_MOESM6_ESM.png

Additional file 6. Maximum-likelihood phylogenetic analysis of full-length BLV-pol gene sequences representing 7 BLV genotypes (G1, G2, G3, G4, G6, G9, and G10) (A); (B) env-based sequences assigned to 10 BLV genotypes (G1, G2, G3, G4, G5, G6, G7, G8, G9, and G10); (C) LTR-based sequences representing 10 BLV genotypes (G1-G10). For all genes and LTR region the Tamura-Nei model and Bootstrap replications (1,000) were applied in MEGA X

12917_2024_4228_MOESM7_ESM.pdf

Additional file 7. Multiple sequence alignment of reverse transcriptase, integrase, envelope and LTR sequences in the context of the specific primers used by different qPCR assays. (A) Multiple sequence alignment of reverse transcriptase (pol gene) sequences in the context of qPCR7, qPCR8, qPCR9, qPCR10 and qPCR11 assay primers. (B) Multiple sequence alignment of integrase (pol gene) sequences in the context of qPCR4 assay primers. (C) Multiple sequence alignment of env gene sequences in the context of ddPCR6. (D) Sequence alignment of LTR region sequences in the context of qPCR3 method primers

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Pluta, A., Jaworski, J.P., Droscha, C. et al. Inter-laboratory comparison of eleven quantitative or digital PCR assays for detection of proviral bovine leukemia virus in blood samples. BMC Vet Res 20 , 381 (2024). https://doi.org/10.1186/s12917-024-04228-z

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Received : 24 November 2023

Accepted : 09 August 2024

Published : 26 August 2024

DOI : https://doi.org/10.1186/s12917-024-04228-z

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