title: Development and Validation of a Highly Accurate Quantitative Real-Time PCR Assay for Diagnosis of Bacterial Vaginosis.
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 orderClostridiales),Megasphaeraphylotype 1 or 2,Lactobacillus iners,Lactobacillus crispatus,Lactobacillus gasseri, andLactobacillus jensenii We generated a logistic regression model that identifiedG. vaginalis,A. vaginae, andMegasphaeraphylotypes 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 ofLactobacillusspp. 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.
authors.affiliation: Femeris Women's Health Research Center, Medical Diagnostic Laboratories, a member of Genesis Biotechnology group, Hamilton, New Jersey, USA dhilbert@femeris.com sgygax@femeris.com.
authors.forename: David W
authors.initials: DW
authors.lastname: Hilbert
authors.affiliation: Femeris Women's Health Research Center, Medical Diagnostic Laboratories, a member of Genesis Biotechnology group, Hamilton, New Jersey, USA.
authors.forename: William L
authors.initials: WL
authors.lastname: Smith
authors.affiliation: Femeris Women's Health Research Center, Medical Diagnostic Laboratories, a member of Genesis Biotechnology group, Hamilton, New Jersey, USA.
authors.forename: Sean G
authors.initials: SG
authors.lastname: Chadwick
authors.affiliation: Femeris Women's Health Research Center, Medical Diagnostic Laboratories, a member of Genesis Biotechnology group, Hamilton, New Jersey, USA.
authors.forename: Geoffrey
authors.initials: G
authors.lastname: Toner
authors.affiliation: Femeris Women's Health Research Center, Medical Diagnostic Laboratories, a member of Genesis Biotechnology group, Hamilton, New Jersey, USA.
authors.forename: Eli
authors.initials: E
authors.lastname: Mordechai
authors.affiliation: Femeris Women's Health Research Center, Medical Diagnostic Laboratories, a member of Genesis Biotechnology group, Hamilton, New Jersey, USA.
authors.forename: Martin E
authors.initials: ME
authors.lastname: Adelson
authors.affiliation: Department of Obstetrics and Gynecology, Wayne State University School of Medicine, Detroit, Michigan, USA.
authors.forename: Tina J
authors.initials: TJ
authors.lastname: Aguin
authors.affiliation: Division of Infectious Diseases, Wayne State University School of Medicine, Detroit, Michigan, USA.
authors.forename: Jack D
authors.initials: JD
authors.lastname: Sobel
authors.affiliation: Femeris Women's Health Research Center, Medical Diagnostic Laboratories, a member of Genesis Biotechnology group, Hamilton, New Jersey, USA dhilbert@femeris.com sgygax@femeris.com.
authors.forename: Scott E
authors.initials: SE
authors.lastname: Gygax
mesh_terms: D000293:Adolescent; D000328:Adult; D000818:Animals; D005260:Female; D006801:Humans; D008137:Longitudinal Studies; D008828:Microbiological Techniques / Q000379:methods*; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014481:United States; D016585:Vaginosis, Bacterial / Q000175:diagnosis*; D055815:Young Adult
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