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1 July 2008

Volume 47, Number 1
Clinical Infectious Diseases 2008;47:33–43
© 2008 by the Infectious Diseases Society of America. All rights reserved.
1058-4838/2008/4701-0006$15.00
DOI: 10.1086/588661
MAJOR ARTICLE

Molecular Quantification of Gardnerella vaginalis and Atopobium vaginae Loads to Predict Bacterial Vaginosis

Jean‐Pierre Menard,1,3

Florence Fenollar,1,2

Mireille Henry,2

Florence Bretelle,3 and

Didier Raoult1,2

1Unité des rickettsies, IFR 48, CNRS UMR 6020, Faculté de Médecine, Université de la Méditerranée, 2Pôle de Maladies Infectieuses, and 3Service de Gynécologie Obstétrique, Marseille, France

Background.Bacterial vaginosis (BV) is a poorly detected public health problem that is associated with preterm delivery and for which no reliable diagnostic tool exists.

Methods.Molecular analysis of 231 vaginal samples, classified by Gram stain–based Nugent score, was used to propose molecular criteria for BV; these criteria were prospectively applied to 56 new samples. A quantitative molecular tool targeting 8 BV‐related microorganisms and a human gene was developed using a specific real‐time polymerase chain reaction assay and serial dilutions of a plasmid suspension. The targeted microorganisms were Gardnerella vaginalis, Lactobacillus species, Mobiluncus curtisii, Mobiluncus mulieris, and Candida albicans (which can be identified by Gram staining), as well as Atopobium vaginae, Mycoplasma hominis, and Ureaplasma urealyticum (which cannot be detected by Gram staining).

Results.With use of the Nugent score, 167 samples were classified as normal, 20 were classified as BV, and 44 were classified as intermediate. Except for U. urealyticum, M. mulieris, and Lactobacillus species, DNA of the tested bacteria was detected more frequently in samples demonstrating BV, but the predictive value of such detection was low. The molecular quantification of A. vaginae (DNA level, 108 copies/mL) and G. vaginalis (DNA level, 109 copies/mL) had the highest predictive value for the diagnosis of BV, with excellent sensitivity (95%), specificity (99%), and positive (95%) and negative (99%) predictive values; 25 (57%) of the samples demonstrating intermediate flora had a BV profile. When applied prospectively, our molecular criteria had total positive and negative predictive values of 96% and 99%, respectively.

Conclusions.We report a highly reproducible, quantitative tool to objectively analyze vaginal flora that uses cutoff values for the concentrations of A. vaginae and G. vaginalis to establish the molecular diagnosis of BV.

Received 20 September 2007; accepted 20 February 2008; electronically published 29 May 2008.

Reprints or correspondence: Dr. Didier Raoult, Unité des rickettsies, IFR 48, CNRS UMR 6020, Faculté de Médecine, Université de la Méditerranée, 27 Blvd. Jean Moulin, 13385 Marseille Cedex 5, France ().

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A. El Khechine, M. Henry, D. Raoult, M. Drancourt. (2009) Detection of Mycobacterium tuberculosis complex organisms in the stools of patients with pulmonary tuberculosis. Microbiology 155:7, 2384-2389
Online publication date: 1-Aug-2009.
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E. De Backer, L. Dubreuil, M. Brauman, J. Acar, M. Vaneechoutte. (2009) In vitro activity of secnidazole against Atopobium vaginae , an anaerobic pathogen involved in bacterial vaginosis. Clinical Microbiology and Infection
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Rebecca M. Brotman and Jacques Ravel. (2008) Editorial Commentary: Ready or Not: The Molecular Diagnosis of Bacterial Vaginosis. Clinical Infectious Diseases 47:1, 44-46
Online publication date: 1-Jul-2008.
Citation-Full Text-PDF Version (158 kB)
Sujatha Srinivasan, David N. Fredricks. (2008) The Human Vaginal Bacterial Biota and Bacterial Vaginosis. Interdisciplinary Perspectives on Infectious Diseases 2008, 1-23
Online publication date: 1-Feb-2008.
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