Comparison of Culture-dependent and Culture-Independent Molecular Methods for Characterisation of Vaginal Microflora.

To date, molecular methods that circumvent the limitations of traditional culture methods have not been used to describe the vaginal microflora in India. Here, we compared culture and culture-independent molecular methods in characterising the vaginal microbiota in Indian women. Culture methods involved traditional cultivation on Rogosa and sheep-blood agar, whereas culture-independent methods bypassed a culturing step by performing broad-range 16S rDNA PCR on DNA isolated directly from vaginal swabs. A total of 13 women were included in the study, of which five were characterised as healthy, two were BV-intermediate and six were BV-positive according to Nugent scoring. Lactobacillus jensenii was detected most frequently when using culture methods. On the other hand, Lactobacillus iners, which was not detected by culture methods, was the most common lactobacillus species detected using cultivation-independent methods. We found little overlap between the species found using cultivation-dependent and cultivation-independent methods. Culture-independent methods were found to be superior in detecting clinically relevant vaginal flora.