Next generation sequencing to examine associations between vaginal washing and vaginal microbiota: A cohort study

Background The association between vaginal washing and HIV risk may be mediated by vaginal washing-associated changes in vaginal microbiota. Methods Data from a cohort of HIV-negative US and Kenyan women enrolled in the Preventing Vaginal Infections trial were analyzed. Vaginal fluid samples and vaginal washing data were collected every 2 months for 12 months. Bacterial relative abundances were measured by broad-range 16S rRNA gene polymerase chain reaction with next generation sequencing. Generalized estimating equations were used to evaluate the association between vaginal washing and i) the Shannon Diversity Index (SDI); and ii) mean change in percent bacterial relative abundances, with application of a 10% false discovery rate (FDR). Results Participants ( N = 111) contributed 93/630 (14.8%) vaginal washing visits. Mean SDI was 0.74 points higher (95% CI 0.35, 1.14; p < 0.001) at washing visits among US participants ( N = 26). Vaginal washing was not associated with SDI in Kenyan participants ( N = 85). There were no associations between vaginal washing and vaginal bacterial relative abundances after applying the FDR. Conclusions The discordant results in Kenyan versus US women suggests the link between vaginal washing and sub-optimal vaginal microbiota may be context specific. Vaginal microbial shifts may not fully explain the association between vaginal washing and HIV acquisition.