Development and Validation of a Semiquantitative, Multitarget PCR Assay for Diagnosis of Bacterial Vaginosis
Open Access
- 1 July 2012
- journal article
- research article
- Published by American Society for Microbiology in Journal of Clinical Microbiology
- Vol. 50 (7), 2321-2329
- https://doi.org/10.1128/jcm.00506-12
Abstract
Quantitative PCR assays were developed for 4 organisms reported previously to be useful positive indicators for the diagnosis of bacterial vaginosis (BV)—Atopobium vaginae, Bacterial Vaginosis-Associated Bacterium 2 (BVAB-2), Gardnerella vaginalis, and Megasphaera-1—and a single organism (Lactobacillus crispatus) that has been implicated as a negative indicator for BV. Vaginal samples (n = 169), classified as positive (n = 108) or negative (n = 61) for BV based on a combination of the Nugent Gram stain score and Amsel clinical criteria, were analyzed for the presence and quantity of each of the marker organisms, and the results were used to construct a semiquantitative, multiplex PCR assay for BV based on detection of 3 positive indicator organisms (A. vaginae, BVAB-2, and Megasphaera-1) and classification of samples using a combinatorial scoring system. The prototype BV PCR assay was then used to analyze the 169-member developmental sample set and, in a prospective, blinded manner, an additional 227 BV-classified vaginal samples (110 BV-positive samples and 117 BV-negative samples). The BV PCR assay demonstrated a sensitivity of 96.7% (202/209), a specificity of 92.2% (153/166), a positive predictive value of 94.0%, and a negative predictive value of 95.6%, with 21 samples (5.3%) classified as indeterminate for BV. This assay provides a reproducible and objective means of evaluating critical components of the vaginal microflora in women with signs and symptoms of vaginitis and is comparable in diagnostic accuracy to the conventional gold standard for diagnosis of BV.Keywords
This publication has 27 references indexed in Scilit:
- 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
- Multiplex Detection of Bacteria Associated with Normal Microbiota and with Bacterial Vaginosis in Vaginal Swabs by Use of Oligonucleotide-Coupled Fluorescent MicrospheresJournal of Clinical Microbiology, 2009
- Changes in Vaginal Bacterial Concentrations with Intravaginal Metronidazole Therapy for Bacterial Vaginosis as Assessed by Quantitative PCRJournal of Clinical Microbiology, 2009
- Quantitative variations in the vaginal bacterial population associated with asymptomatic infections: a real-time polymerase chain reaction studyEuropean Journal of Clinical Microbiology & Infectious Diseases, 2008
- Molecular Quantification ofGardnerella vaginalisandAtopobium vaginaeLoads to Predict Bacterial VaginosisClinical Infectious Diseases, 2008
- Association between Lactobacillus species and bacterial vaginosis-related bacteria, and bacterial vaginosis scores in pregnant Japanese womenBMC Infectious Diseases, 2007
- Targeted PCR for Detection of Vaginal Bacteria Associated with Bacterial VaginosisJournal of Clinical Microbiology, 2007
- Cultivation-Independent Analysis of Changes in Bacterial Vaginosis Flora Following Metronidazole TreatmentJournal of Clinical Microbiology, 2007
- Prevalence of Bacterial Vaginosis and Vaginal Flora Changes in Peri- and Postmenopausal WomenJournal of Clinical Microbiology, 2002