Peer-Reviewed Study Confirms Accuracy of Evvy’s Vaginal Microbiome Test

The vaginal microbiome is finally getting the attention it deserves, but let’s face it, it’s still a relatively new field of research. Our mission at Evvy has always been to close the gender health gap by discovering and leveraging overlooked female biomarkers — starting with the vaginal microbiome — and we’ve always prided ourselves in being a science-forward company at the forefront of research and innovation. 

That’s why we are excited to announce that our vaginal microbiome test validation process has been published in the peer-reviewed journal Diagnostics. 

The Evvy Vaginal Health Test is now the only vaginal microbiome test in the world to be both peer-reviewed and analytically validated by CLIA, CAP and CLEP. It’s an important step forward for the Evvy community and the scientific community as a whole.

What does it mean that Evvy is “peer-reviewed” and “independently validated”? 

To have a paper peer-reviewed means that it was submitted to a journal, which then sent it to three experts in the field. These experts not only read through the paper but also critiqued every detail of it. We, the authors of the paper, received the peer-reviewed comments, re-analyzed the paper, and addressed all of their concerns before submitting it for final publication. This rigorous review process is essential for the scientific community to trust the validity of publications. We also hope it sets a precedent for other vaginal microbiome testing companies to aim for the same standards.

As for CLIA, CAP, and CLEP certification, what does that mean? It means that our test protocol has been scrutinized by not one but three different governmental agencies: 

• The Clinical Laboratory Improvement Amendments (CLIA)
• The College of American Pathologists (CAP)
• And the New York-based Clinical Laboratory Evaluation Program (CLEP). 

In addition to being scrutinized by these regulatory agencies, we also wanted scientists to analyze our methods, which is why we published a peer-reviewed paper describing our test validation. 

Since the vaginal microbiome has become more and more popular, several brands offer testing methods — but not all vaginal microbiome tests are created equal. Everything from the sequencing technology used to the validity of the lab affects how accurate a vaginal microbiome test is.

We’ve been working hard behind the scenes to publish our validation process and show Evvy customers that our test is the most reliable out there. At the very least, it’s a guarantee that with Evvy, you’re in safe hands—and you get what you paid for!

Peer-reviewed results confirm the accuracy of Evvy’s Vaginal Health Test

From this validation paper, we know that the Evvy test:

• Has a sensitivity of 93.1%, a specificity of 90%, a negative predictive value of 93.4%, and a positive predictive value of 89.6% based on the ability to detect over 160 pure culture isolates (samples of a particular bacteria or fungi). This metric tells you how much you can trust the test. When we compare experiments using pure culture isolates we can detect those specific organisms more than 90% of the time, and our classification is highly accurate. Meaning the test is highly sensitive and specific. 
• Has a limit of detection of 3000–1640 CFU/mL for bacteria and 2400 CFU/mL for fungi. This metric tells us how many organisms must be present in a sample for the test to detect it (aka our limit of detection). Our test shows that for fungi and different types of bacteria (Gram-positive and Gram-negative), there needs to be about 1,600 to 3000 cells in a milliliter of liquid (CFU/ml). They may sound like a lot, but a normal vaginal swab sample would have anywhere from 10⁷ to 10⁹ CFU/ml (that’s 10,000,000 to 1,000,000,000 cells). This means that even if your microbiome had 10,000 to 100,000 fewer cells than the normal range, we would be able to detect it!

Our sequencing technology

Not all sequencing methods can provide species-level identification for all the organisms in the vagina, but the Evvy test can. Thanks to the type of sequencing we use (metagenomics) and our vast dataset, we provide you with more specific identification of the organisms you care about, like Gardnerella, Prevotella, and Lactobacillus.

Gardnerella

The science of the vaginal microbiome is advancing rapidly, and one example is Gardnerella. Up until about 2019, only one species of Gardnerella was identified, since then four species have been named and six others have yet to be named. Meaning that there is still a lot to be learned. 

When we developed the Evvy test we wanted to be sure that our test detects all the variation in this important vaginal organism, so we worked hard to make sure our test detects as many Gardnerella species as possible. 

Fun fact about Gardnerella, the reason we didn't know so many different species existed until recently is because they have similar 16S genes, you need to be able to look at the whole genome to tell the difference between organisms. So while PCR and 16S sequencing might detect only one or two of the species, the Evvy test can detect all of them. 

Prevotella

Another organism that is very important for vaginal health is Prevotella. This bacterium is thought to be one of the main drivers of vaginal odor and discomfort. Unfortunately, Prevotella species are notoriously difficult to study, as they're fastidious bacteria and require very specific growing conditions. As a result, there are very few Prevotella genomes available. So most reference databases don’t contain many Prevotella reference genomes. 

While developing Evvy’s reference database, we noticed this gap and decided to fill it ourselves. Using a bioinformatics approach called metagenome-assembled genomes (MAG), we were able to almost triple the number of Prevotella species in our database. This means that when detecting Prevotella, we can be as accurate as possible. 

We have done this same process for other BV-associated organisms Fannyhessae (previously Atopobium), Megasphaera, Porphyromonas, Snethia, Ureaplasma, and many more. 

How Evvy users are contributing to women’s health research

One of the first questions we ask you when you take a test is if you want to opt-in to being included in research. And the majority (97.5%) of you say yes! 

This means a lot, that so many of you want to help contribute to bridging the gender health gap, and we take this responsibility very seriously. As of 2024, we have published in journals and presented preliminary data on multiple studies at national and international conferences. Here are a few of the things we have learned so far:

1. A review on the mental health impact of urogenital infections, published in the Journal of Women’s Health.
2. Different Gardnerella species have different metabolism which means that different Gardnerella species will have different effects on the vaginal microbiome. Published in mSystems a journal for the American Society of Microbiology.
3. The presence of aerobes (oxygen-loving organisms) in vaginitis patients results in a greater number and severity of symptoms. Presented at the Infectious Disease Society of ObGyns (IDSOG) conference in 2023. 
4. There are differences in the pathogenicity genes in the genomes of Gardnerella. Presented at the Infectious Disease Society of ObGyn (IDSOG) conference in 2023.  
5. Women who report pain with sex have different profiles based on age. Menopausal women are more diverse than reproductive-aged women. Presented at the International Society of Women’s Sexual Health (ISSWSH) Conference in 2024. 
6. Candida albicans can be present in women with high relative abundance of Lactobacillus with or without symptoms. Presented at the International Society for the Study of Vulvovaginal Diseases (ISSVD) Conference in 2024. 
7. Vaginal odor is associated with increased abundance of Prevotella, Megasphaera, Porphyromonas, and Finegoldia. Presented at the Infectious Disease Society of ObGyn (IDSOG) conference in 2024. 

Evidence-based information

Evvy’s science-forward approach goes beyond our product. We want every Evvy user to have access to all the information they need to make informed decisions about their health. However, the field of science can be complex, and thanks to the gender health gap, much remains unknown about female reproductive health. This is why we have the #AskEvvy blog to help explain the science. We take this blog very seriously; each article undergoes both scientific and medical reviews before publication.

We also don’t ever make claims that we can’t back up. We provide citations for our sources and offer only evidence-based information. We include a link to the citation whenever we mention an association or a specific bacterium. Our goal is to ensure that the information we provide is as accurate as possible. 

We’re not done yet

Everything that was described above is the best we can do right now. But we learn more every day, and science is always progressing. As we learn more, we pass that along to you. 

That means that we are continuously improving our lab processes, reference databases, and updating our information here on the blog and in the product. We do our best to provide you with the most accurate information possible. And in return,  we hope to continuously learn together.