A novel Gardnerella, Prevotella, and Lactobacillus standard that improves accuracy in quantifying bacterial burden in vaginal microbial communities

Authors

Jacob H. Elnaggar, LSU Health Sciences Center - New OrleansFollow
Caleb M. Ardizzone, LSU Health Sciences Center - New OrleansFollow
Nuno Cerca, Universidade do Minho
Evelyn Toh, Indiana University School of Medicine
Paweł Łaniewski, University of Arizona College of Medicine – Phoenix
Rebecca A. Lillis, LSU Health Sciences Center - New OrleansFollow
Melissa M. Herbst-Kralovetz, University of Arizona College of Medicine – Phoenix
Alison J. Quayle, LSU Health Sciences Center - New OrleansFollow
Christina A. Muzny, The University of Alabama at Birmingham
Christopher M. Taylor, LSU Health Sciences Center - New OrleansFollow

Document Type

Article

Publication Date

6-19-2023

Publication Title

Frontiers in Cellular and Infection Microbiology

Abstract

Bacterial vaginosis (BV) is the most common vaginal dysbiosis. In this condition, a polymicrobial biofilm develops on vaginal epithelial cells. Accurately quantifying the bacterial burden of the BV biofilm is necessary to further our understanding of BV pathogenesis. Historically, the standard for calculating total bacterial burden of the BV biofilm has been based on quantifying Escherichia coli 16S rRNA gene copy number. However, E. coli is improper for measuring the bacterial burden of this unique micro-environment. Here, we propose a novel qPCR standard to quantify bacterial burden in vaginal microbial communities, from an optimal state to a mature BV biofilm. These standards consist of different combinations of vaginal bacteria including three common BV-associated bacteria (BVAB) Gardnerella spp. (G), Prevotella spp. (P), and Fannyhessea spp. (F) and commensal Lactobacillus spp. (L) using the 16S rRNA gene (G:P:F:L, G:P:F, G:P:L and 1G:9L). We compared these standards to the traditional E. coli (E) reference standard using known quantities of mock vaginal communities and 16 vaginal samples from women. The E standard significantly underestimated the copy numbers of the mock communities, and this underestimation was significantly greater at lower copy numbers of these communities. The G:P:L standard was the most accurate across all mock communities and when compared to other mixed vaginal standards. Mixed vaginal standards were further validated with vaginal samples. This new G:P:L standard can be used in BV pathogenesis research to enhance reproducibility and reliability in quantitative measurements of BVAB, spanning from the optimal to non-optimal (including BV) vaginal microbiota.

PubMed ID

37404722

Volume

13

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Recommended Citation

Elnaggar, Jacob H.; Ardizzone, Caleb M.; Cerca, Nuno; Toh, Evelyn; Łaniewski, Paweł; Lillis, Rebecca A.; Herbst-Kralovetz, Melissa M.; Quayle, Alison J.; Muzny, Christina A.; and Taylor, Christopher M., "A novel Gardnerella, Prevotella, and Lactobacillus standard that improves accuracy in quantifying bacterial burden in vaginal microbial communities" (2023). School of Graduate Studies Faculty Publications. 128.
https://digitalscholar.lsuhsc.edu/sogs_facpubs/128
10.3389/fcimb.2023.1198113