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

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 c...

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Autores principales: 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., Taylor, Christopher M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10315654/
https://www.ncbi.nlm.nih.gov/pubmed/37404722
http://dx.doi.org/10.3389/fcimb.2023.1198113
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author 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.
Taylor, Christopher M.
author_facet 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.
Taylor, Christopher M.
author_sort Elnaggar, Jacob H.
collection PubMed
description 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.
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spelling pubmed-103156542023-07-04 A novel Gardnerella, Prevotella, and Lactobacillus standard that improves accuracy in quantifying bacterial burden in vaginal microbial communities 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. Taylor, Christopher M. Front Cell Infect Microbiol Cellular and Infection Microbiology 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. Frontiers Media S.A. 2023-06-19 /pmc/articles/PMC10315654/ /pubmed/37404722 http://dx.doi.org/10.3389/fcimb.2023.1198113 Text en Copyright © 2023 Elnaggar, Ardizzone, Cerca, Toh, Łaniewski, Lillis, Herbst-Kralovetz, Quayle, Muzny and Taylor https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cellular and Infection Microbiology
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.
Taylor, Christopher M.
A novel Gardnerella, Prevotella, and Lactobacillus standard that improves accuracy in quantifying bacterial burden in vaginal microbial communities
title A novel Gardnerella, Prevotella, and Lactobacillus standard that improves accuracy in quantifying bacterial burden in vaginal microbial communities
title_full A novel Gardnerella, Prevotella, and Lactobacillus standard that improves accuracy in quantifying bacterial burden in vaginal microbial communities
title_fullStr A novel Gardnerella, Prevotella, and Lactobacillus standard that improves accuracy in quantifying bacterial burden in vaginal microbial communities
title_full_unstemmed A novel Gardnerella, Prevotella, and Lactobacillus standard that improves accuracy in quantifying bacterial burden in vaginal microbial communities
title_short A novel Gardnerella, Prevotella, and Lactobacillus standard that improves accuracy in quantifying bacterial burden in vaginal microbial communities
title_sort novel gardnerella, prevotella, and lactobacillus standard that improves accuracy in quantifying bacterial burden in vaginal microbial communities
topic Cellular and Infection Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10315654/
https://www.ncbi.nlm.nih.gov/pubmed/37404722
http://dx.doi.org/10.3389/fcimb.2023.1198113
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