Development and Validation of a Highly Accurate Quantitative Real-Time PCR Assay for Diagnosis of Bacterial Vaginosis
Open Access
- 1 April 2016
- journal article
- research article
- Published by American Society for Microbiology in Journal of Clinical Microbiology
- Vol. 54 (4), 1017-1024
- https://doi.org/10.1128/jcm.03104-15
Abstract
Bacterial vaginosis (BV) is the most common gynecological infection in the United States. Diagnosis based on Amsel's criteria can be challenging and can be aided by laboratory-based testing. A standard method for diagnosis in research studies is enumeration of bacterial morphotypes of a Gram-stained vaginal smear (i.e., Nugent scoring). However, this technique is subjective, requires specialized training, and is not widely available. Therefore, a highly accurate molecular assay for the diagnosis of BV would be of great utility. We analyzed 385 vaginal specimens collected prospectively from subjects who were evaluated for BV by clinical signs and Nugent scoring. We analyzed quantitative real-time PCR (qPCR) assays on DNA extracted from these specimens to quantify nine organisms associated with vaginal health or disease: Gardnerella vaginalis, Atopobium vaginae, BV-associated bacteria 2 (BVAB2, an uncultured member of the order Clostridiales), Megasphaera phylotype 1 or 2, Lactobacillus iners, Lactobacillus crispatus, Lactobacillus gasseri, and Lactobacillus jensenii. We generated a logistic regression model that identified G. vaginalis, A. vaginae, and Megasphaera phylotypes 1 and 2 as the organisms for which quantification provided the most accurate diagnosis of symptomatic BV, as defined by Amsel's criteria and Nugent scoring, with 92% sensitivity, 95% specificity, 94% positive predictive value, and 94% negative predictive value. The inclusion of Lactobacillus spp. did not contribute sufficiently to the quantitative model for symptomatic BV detection. This molecular assay is a highly accurate laboratory tool to assist in the diagnosis of symptomatic BV.Keywords
This publication has 39 references indexed in Scilit:
- Development and Validation of a Semiquantitative, Multitarget PCR Assay for Diagnosis of Bacterial VaginosisJournal of Clinical Microbiology, 2012
- Vaginal microbiome of reproductive-age womenProceedings of the National Academy of Sciences, 2010
- Quantitative PCR Assessments of Bacterial Species in Women with and without Bacterial VaginosisJournal of Clinical Microbiology, 2010
- Longitudinal analysis of the vaginal microflora in pregnancy suggests that L. crispatus promotes the stability of the normal vaginal microflora and that L. gasseri and/or L. iners are more conducive to the occurrence of abnormal vaginal microfloraBMC Microbiology, 2009
- Molecular Quantification ofGardnerella vaginalisandAtopobium vaginaeLoads to Predict Bacterial VaginosisClinical Infectious Diseases, 2008
- Targeted PCR for Detection of Vaginal Bacteria Associated with Bacterial VaginosisJournal of Clinical Microbiology, 2007
- A randomized trial of metronidazole in asymptomatic bacterial vaginosis to prevent the acquisition of sexually transmitted diseasesAmerican Journal of Obstetrics and Gynecology, 2007
- Culture-independent analysis of vaginal microflora: The unrecognized association of Atopobium vaginae with bacterial vaginosisAmerican Journal of Obstetrics and Gynecology, 2004
- Predictive value of the clinical diagnosis of lower genital tract infection in womenAmerican Journal of Obstetrics and Gynecology, 2004
- Anaerobic Bacteria in Nonspecific VaginitisNew England Journal of Medicine, 1980