Objectives
Gardnerella vaginalis (GV) is highly prevalent in women with abnormal vaginal flora (Nugent score 4-6), yet have a preponderance of lactobacilli. Recent literature has suggested that specific clades (groupings based on genomic similarities) of GV may have different virulence factors with some more likely to be associated with bacterial vaginosis (BV) (Nugent score 7-10). The objective of this study was to conduct a prospective, longitudinal study of women with GV but no evidence of BV to determine if there were strain differences associated with the development of BV.
Methods
Women without evidence of BV by Nugent score were followed longitudinally for up to 8 weeks or the development of BV. They provided weekly vaginal slides to be interpreted by Nugent criteria, and vaginal swab specimens for GV PCR. For GV PCR, DNA was extracted from 100 μL of vaginal swab wash using a two-step digest (50 U/μL Ready-Lyse lysozyme at 37°C overnight, followed by 1 hour at 56°C with 120 nG/μL Proteinase K and 60 mM DTT. The digest was used as the source material for a standard column-based extraction method. The presence and stratification of GV clades in specimens was determined using the procedure outlined by Balashov, et. al, 2014 adapted for amplification on a Roche LightCycler Z480. Statistical analyses were conducted using SAS v9.4 and OpenEpi v2015/05/04.
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