Rapid Quantitative PCR Assay for the Detection of the Three Vaginal Pathogens Candida, Gardnerella and Atopobium as well as the Commensal Lactobacillus Genera

Stephanie Bornes; Olivier Camarès; Marylise Paquet-Gachinat; Philippe Veisseire; Jacques Ravel; Caroline Dausset; Adrien Nivoliez

doi:doi:10.11648/j.ijmb.20210603.12

Rapid Quantitative PCR Assay for the Detection of the Three Vaginal Pathogens Candida, Gardnerella and Atopobium as well as the Commensal Lactobacillus Genera

Stephanie Bornes, Olivier Camarès, Marylise Paquet-Gachinat, Philippe Veisseire, Jacques Ravel, Caroline Dausset, Adrien Nivoliez

Published in International Journal of Microbiology and Biotechnology (Volume 6, Issue 3)

Received: 12 July 2021 Accepted: 27 July 2021 Published: 4 August 2021

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Abstract

The vaginal microbiota balance is quite fragile and susceptible to the development of vaginosis and candidiasis. The current diagnostic method for bacterial vaginosis relies on the evaluation of different bacterial morphotypes using the Nugent score. This method is only partially in correlation with a DNA sequencing-based diagnostic or Amsel criteria used by clinicians, suggesting the need for new molecular approaches dedicated to the diagnosis of BV. The objective of this study was to develop and validate a quantitative polymerase chain reaction (qPCR) assay for the specific and rapid detection of three vaginal pathogens, i.e. Candida, Gardnerella and Atopobium and the commensal Lactobacillus genera. For this purpose, four oligonucleotide primer pairs were designed and tested to obtain optimal amplification of the four target genera. The qPCR assay was also tested on the non-target genera and on human DNA. The designed primers allowed specific amplification of the target organisms in vitro and in clinical vaginal samples. The qPCR assay designed in this study is effective to specifically detect these genera in clinical samples as a molecular technique complementary to the Nugent score. It can be used in epidemiological studies for understanding the role of these pathogens and to follow their abundance in the microbiota in disease processes such as the development of vulvovaginal candidiasis and bacterial vaginosis.

Published in	International Journal of Microbiology and Biotechnology (Volume 6, Issue 3)
DOI	10.11648/j.ijmb.20210603.12
Page(s)	71-77
Creative Commons	This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.
Copyright	Copyright © The Author(s), 2021. Published by Science Publishing Group

Keywords

Lactobacillus, Atopobium, Gardnerella, Candida, Quantitative PCR, Vaginal Infections

References

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[9]	Boyd Tressler A, Markwei M, Fortin C, Yao M, Procop GW, Soper DE, Goje O. (2021). Risks for Recurrent Vulvovaginal Candidiasis Caused by Non-Albicans Candida Versus Candida Albicans. J Womens Health (Larchmt).
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APA Style

Stephanie Bornes, Olivier Camarès, Marylise Paquet-Gachinat, Philippe Veisseire, Jacques Ravel, et al. (2021). Rapid Quantitative PCR Assay for the Detection of the Three Vaginal Pathogens Candida, Gardnerella and Atopobium as well as the Commensal Lactobacillus Genera. International Journal of Microbiology and Biotechnology, 6(3), 71-77. https://doi.org/10.11648/j.ijmb.20210603.12

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ACS Style

Stephanie Bornes; Olivier Camarès; Marylise Paquet-Gachinat; Philippe Veisseire; Jacques Ravel, et al. Rapid Quantitative PCR Assay for the Detection of the Three Vaginal Pathogens Candida, Gardnerella and Atopobium as well as the Commensal Lactobacillus Genera. Int. J. Microbiol. Biotechnol. 2021, 6(3), 71-77. doi: 10.11648/j.ijmb.20210603.12

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AMA Style

Stephanie Bornes, Olivier Camarès, Marylise Paquet-Gachinat, Philippe Veisseire, Jacques Ravel, et al. Rapid Quantitative PCR Assay for the Detection of the Three Vaginal Pathogens Candida, Gardnerella and Atopobium as well as the Commensal Lactobacillus Genera. Int J Microbiol Biotechnol. 2021;6(3):71-77. doi: 10.11648/j.ijmb.20210603.12

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@article{10.11648/j.ijmb.20210603.12,
  author = {Stephanie Bornes and Olivier Camarès and Marylise Paquet-Gachinat and Philippe Veisseire and Jacques Ravel and Caroline Dausset and Adrien Nivoliez},
  title = {Rapid Quantitative PCR Assay for the Detection of the Three Vaginal Pathogens Candida, Gardnerella and Atopobium as well as the Commensal Lactobacillus Genera},
  journal = {International Journal of Microbiology and Biotechnology},
  volume = {6},
  number = {3},
  pages = {71-77},
  doi = {10.11648/j.ijmb.20210603.12},
  url = {https://doi.org/10.11648/j.ijmb.20210603.12},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijmb.20210603.12},
  abstract = {The vaginal microbiota balance is quite fragile and susceptible to the development of vaginosis and candidiasis. The current diagnostic method for bacterial vaginosis relies on the evaluation of different bacterial morphotypes using the Nugent score. This method is only partially in correlation with a DNA sequencing-based diagnostic or Amsel criteria used by clinicians, suggesting the need for new molecular approaches dedicated to the diagnosis of BV. The objective of this study was to develop and validate a quantitative polymerase chain reaction (qPCR) assay for the specific and rapid detection of three vaginal pathogens, i.e. Candida, Gardnerella and Atopobium and the commensal Lactobacillus genera. For this purpose, four oligonucleotide primer pairs were designed and tested to obtain optimal amplification of the four target genera. The qPCR assay was also tested on the non-target genera and on human DNA. The designed primers allowed specific amplification of the target organisms in vitro and in clinical vaginal samples. The qPCR assay designed in this study is effective to specifically detect these genera in clinical samples as a molecular technique complementary to the Nugent score. It can be used in epidemiological studies for understanding the role of these pathogens and to follow their abundance in the microbiota in disease processes such as the development of vulvovaginal candidiasis and bacterial vaginosis.},
 year = {2021}
}

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TY  - JOUR
T1  - Rapid Quantitative PCR Assay for the Detection of the Three Vaginal Pathogens Candida, Gardnerella and Atopobium as well as the Commensal Lactobacillus Genera
AU  - Stephanie Bornes
AU  - Olivier Camarès
AU  - Marylise Paquet-Gachinat
AU  - Philippe Veisseire
AU  - Jacques Ravel
AU  - Caroline Dausset
AU  - Adrien Nivoliez
Y1  - 2021/08/04
PY  - 2021
N1  - https://doi.org/10.11648/j.ijmb.20210603.12
DO  - 10.11648/j.ijmb.20210603.12
T2  - International Journal of Microbiology and Biotechnology
JF  - International Journal of Microbiology and Biotechnology
JO  - International Journal of Microbiology and Biotechnology
SP  - 71
EP  - 77
PB  - Science Publishing Group
SN  - 2578-9686
UR  - https://doi.org/10.11648/j.ijmb.20210603.12
AB  - The vaginal microbiota balance is quite fragile and susceptible to the development of vaginosis and candidiasis. The current diagnostic method for bacterial vaginosis relies on the evaluation of different bacterial morphotypes using the Nugent score. This method is only partially in correlation with a DNA sequencing-based diagnostic or Amsel criteria used by clinicians, suggesting the need for new molecular approaches dedicated to the diagnosis of BV. The objective of this study was to develop and validate a quantitative polymerase chain reaction (qPCR) assay for the specific and rapid detection of three vaginal pathogens, i.e. Candida, Gardnerella and Atopobium and the commensal Lactobacillus genera. For this purpose, four oligonucleotide primer pairs were designed and tested to obtain optimal amplification of the four target genera. The qPCR assay was also tested on the non-target genera and on human DNA. The designed primers allowed specific amplification of the target organisms in vitro and in clinical vaginal samples. The qPCR assay designed in this study is effective to specifically detect these genera in clinical samples as a molecular technique complementary to the Nugent score. It can be used in epidemiological studies for understanding the role of these pathogens and to follow their abundance in the microbiota in disease processes such as the development of vulvovaginal candidiasis and bacterial vaginosis.
VL  - 6
IS  - 3
ER  -

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Author Information

Stephanie Bornes

University of Clermont Auvergne, INRAE, VetAgro Sup, UMRF, Aurillac, France
Olivier Camarès

University of Clermont Auvergne, INRAE, VetAgro Sup, UMRF, Aurillac, France
Marylise Paquet-Gachinat

University of Clermont Auvergne, INRAE, VetAgro Sup, UMRF, Aurillac, France
Philippe Veisseire

University of Clermont Auvergne, INRAE, VetAgro Sup, UMRF, Aurillac, France
Jacques Ravel

Institute for Genome Sciences, University of Maryland School of Medicine, Baltimore, USA
Caroline Dausset

Department of Research and Development, Biose Industry, Arpajon-Sur-Cère, France
Adrien Nivoliez

Department of Research and Development, Biose Industry, Arpajon-Sur-Cère, France

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APA Style

Stephanie Bornes, Olivier Camarès, Marylise Paquet-Gachinat, Philippe Veisseire, Jacques Ravel, et al. (2021). Rapid Quantitative PCR Assay for the Detection of the Three Vaginal Pathogens Candida, Gardnerella and Atopobium as well as the Commensal Lactobacillus Genera. International Journal of Microbiology and Biotechnology, 6(3), 71-77. https://doi.org/10.11648/j.ijmb.20210603.12

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ACS Style

Stephanie Bornes; Olivier Camarès; Marylise Paquet-Gachinat; Philippe Veisseire; Jacques Ravel, et al. Rapid Quantitative PCR Assay for the Detection of the Three Vaginal Pathogens Candida, Gardnerella and Atopobium as well as the Commensal Lactobacillus Genera. Int. J. Microbiol. Biotechnol. 2021, 6(3), 71-77. doi: 10.11648/j.ijmb.20210603.12

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AMA Style

Stephanie Bornes, Olivier Camarès, Marylise Paquet-Gachinat, Philippe Veisseire, Jacques Ravel, et al. Rapid Quantitative PCR Assay for the Detection of the Three Vaginal Pathogens Candida, Gardnerella and Atopobium as well as the Commensal Lactobacillus Genera. Int J Microbiol Biotechnol. 2021;6(3):71-77. doi: 10.11648/j.ijmb.20210603.12

Copy | Download

@article{10.11648/j.ijmb.20210603.12,
  author = {Stephanie Bornes and Olivier Camarès and Marylise Paquet-Gachinat and Philippe Veisseire and Jacques Ravel and Caroline Dausset and Adrien Nivoliez},
  title = {Rapid Quantitative PCR Assay for the Detection of the Three Vaginal Pathogens Candida, Gardnerella and Atopobium as well as the Commensal Lactobacillus Genera},
  journal = {International Journal of Microbiology and Biotechnology},
  volume = {6},
  number = {3},
  pages = {71-77},
  doi = {10.11648/j.ijmb.20210603.12},
  url = {https://doi.org/10.11648/j.ijmb.20210603.12},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijmb.20210603.12},
  abstract = {The vaginal microbiota balance is quite fragile and susceptible to the development of vaginosis and candidiasis. The current diagnostic method for bacterial vaginosis relies on the evaluation of different bacterial morphotypes using the Nugent score. This method is only partially in correlation with a DNA sequencing-based diagnostic or Amsel criteria used by clinicians, suggesting the need for new molecular approaches dedicated to the diagnosis of BV. The objective of this study was to develop and validate a quantitative polymerase chain reaction (qPCR) assay for the specific and rapid detection of three vaginal pathogens, i.e. Candida, Gardnerella and Atopobium and the commensal Lactobacillus genera. For this purpose, four oligonucleotide primer pairs were designed and tested to obtain optimal amplification of the four target genera. The qPCR assay was also tested on the non-target genera and on human DNA. The designed primers allowed specific amplification of the target organisms in vitro and in clinical vaginal samples. The qPCR assay designed in this study is effective to specifically detect these genera in clinical samples as a molecular technique complementary to the Nugent score. It can be used in epidemiological studies for understanding the role of these pathogens and to follow their abundance in the microbiota in disease processes such as the development of vulvovaginal candidiasis and bacterial vaginosis.},
 year = {2021}
}

Copy | Download

TY  - JOUR
T1  - Rapid Quantitative PCR Assay for the Detection of the Three Vaginal Pathogens Candida, Gardnerella and Atopobium as well as the Commensal Lactobacillus Genera
AU  - Stephanie Bornes
AU  - Olivier Camarès
AU  - Marylise Paquet-Gachinat
AU  - Philippe Veisseire
AU  - Jacques Ravel
AU  - Caroline Dausset
AU  - Adrien Nivoliez
Y1  - 2021/08/04
PY  - 2021
N1  - https://doi.org/10.11648/j.ijmb.20210603.12
DO  - 10.11648/j.ijmb.20210603.12
T2  - International Journal of Microbiology and Biotechnology
JF  - International Journal of Microbiology and Biotechnology
JO  - International Journal of Microbiology and Biotechnology
SP  - 71
EP  - 77
PB  - Science Publishing Group
SN  - 2578-9686
UR  - https://doi.org/10.11648/j.ijmb.20210603.12
AB  - The vaginal microbiota balance is quite fragile and susceptible to the development of vaginosis and candidiasis. The current diagnostic method for bacterial vaginosis relies on the evaluation of different bacterial morphotypes using the Nugent score. This method is only partially in correlation with a DNA sequencing-based diagnostic or Amsel criteria used by clinicians, suggesting the need for new molecular approaches dedicated to the diagnosis of BV. The objective of this study was to develop and validate a quantitative polymerase chain reaction (qPCR) assay for the specific and rapid detection of three vaginal pathogens, i.e. Candida, Gardnerella and Atopobium and the commensal Lactobacillus genera. For this purpose, four oligonucleotide primer pairs were designed and tested to obtain optimal amplification of the four target genera. The qPCR assay was also tested on the non-target genera and on human DNA. The designed primers allowed specific amplification of the target organisms in vitro and in clinical vaginal samples. The qPCR assay designed in this study is effective to specifically detect these genera in clinical samples as a molecular technique complementary to the Nugent score. It can be used in epidemiological studies for understanding the role of these pathogens and to follow their abundance in the microbiota in disease processes such as the development of vulvovaginal candidiasis and bacterial vaginosis.
VL  - 6
IS  - 3
ER  -

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