A novel Gardnerella, Prevotella, and Lactobacillus standard that improves accuracy in quantifying bacterial burden in vaginal microbial communities
Author
Elnaggar, J.H.Ardizzone, C.M.
Cerca, N.
Toh, E.
Łaniewski, P.
Lillis, R.A.
Herbst-Kralovetz, M.M.
Quayle, A.J.
Muzny, C.A.
Taylor, C.M.
Affiliation
Department of Basic Medical Sciences, College of Medicine-Phoenix, University of ArizonaIssue Date
2023-06-18Keywords
bacterial burdenbacterial vaginosis
biofilm
Gardnerella
Lactobacillus
Prevotella
qPCR standard
vaginal microbiome
Metadata
Show full item recordPublisher
Frontiers Media SACitation
Elnaggar JH, Ardizzone CM, Cerca N, Toh E, Łaniewski P, Lillis RA, Herbst-Kralovetz MM, Quayle AJ, Muzny CA and Taylor CM (2023) A novel Gardnerella, Prevotella, and Lactobacillus standard that improves accuracy in quantifying bacterial burden in vaginal microbial communities. Front. Cell. Infect. Microbiol. 13:1198113. doi: 10.3389/fcimb.2023.1198113Rights
© 2023 Elnaggar, Ardizzone, Cerca, Toh, Łaniewski, Lillis, Herbst-Kralovetz, Quayle, Muzny and Taylor. This is an open-access article distributed under the terms of the Creative Commons Attribution License.Collection Information
This item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at repository@u.library.arizona.edu.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. Copyright © 2023 Elnaggar, Ardizzone, Cerca, Toh, Łaniewski, Lillis, Herbst-Kralovetz, Quayle, Muzny and Taylor.Note
Open access journalISSN
2235-2988PubMed ID
37404722Version
Final Published Versionae974a485f413a2113503eed53cd6c53
10.3389/fcimb.2023.1198113
Scopus Count
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Except where otherwise noted, this item's license is described as © 2023 Elnaggar, Ardizzone, Cerca, Toh, Łaniewski, Lillis, Herbst-Kralovetz, Quayle, Muzny and Taylor. This is an open-access article distributed under the terms of the Creative Commons Attribution License.

