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Background  To obtain more detailed understanding of the causes of disturbance of the vaginal microflora (VMF), a longitudinal study was carried out for 17 women during two menstrual cycles.    Methods  Vaginal swabs were obtained daily from 17 non-pregnant, menarchal volunteers. For each woman, Gram stains were scored, the quantitative changes of 5 key vaginal species,  i.e. Atopobium vaginae ,  Lactobacillus crispatus ,  L. iners , (sialidase positive)  Gardnerella vaginalis  and  Prevotella bivia  were quantified with qPCR and hydrogen-peroxide production was assessed on TMB+ agar.    Results  Women could be divided in 9 subjects with predominantly normal VMF (grades Ia, Ib and Iab, group N) and 8 with predominantly disturbed VMF (grades I-like, II, III and IV, group D).  VMF was variable between women, but overall stable for most of the women. Menses were the strongest disturbing factor of the VMF.    L. crispatus  was present at log7–9 cells/ml in grade Ia, Iab and II VMF, but concentrations declined 100-fold during menses.  L. crispatus  below log7 cells/ml corresponded with poor H 2 O 2 -production.  L. iner s was present at log 10 cells/ml in grade Ib, II and III VMF. Sialidase negative  G. vaginalis  strains (average log5 cells/ml) were detected in grade I, I-like and IV VMF. In grade II VMF, predominantly a mixture of both sialidase negative and positive  G. vaginalis  strains (average log9 cells/ml) were present, and predominantly sialidase positive strains in grade III VMF. The presence of  A. vaginae  (average log9 cells/ml) coincided with grade II and III VMF.  P. bivia  (log4–8 cells/ml) was mostly present in grade III vaginal microflora.    L. iners ,  G. vaginalis ,  A. vaginae  and  P. bivia  all increased around menses for group N women, and as such  L. iners  was considered a member of disturbed VMF.    Conclusions  This qPCR-based study confirms largely the results of previous culture-based, microscopy-based and pyrosequencing-based studies.

Longitudinal qPCR Study of the Dynamics of L. crispatus, L. iners, A. vaginae, (Sialidase Positive) G. vaginalis, and P. bivia in the Vagina