Comparison of real-time quantitative PCR and 16S rDNA sequencing for detection of female vaginal microbiome / 上海交通大学学报（医学版）

ABSTRACT

Objective·To compare the differences in the composition of female vaginal flora by real-time quantitative PCR and 16S rDNA sequencing.Methods·Forty-nine healthy reproductive women less than 45 years old were selected.Specimens were collected from posterior fornix.DNA were extracted and the microbiome were ananlyzed by both 16S rDNA V1V2 region sequencing and qPCR of 22 selected genes.The results detected by two methods were compared.Results·According to the classification standard of vaginal community state type (CSTs),qPCR analysis showed that 35 out of 49 samples were dominated by Lactobacillus species,among them,type Ⅰ (Lactobacillus crispatus,9),type Ⅲ (Lactobacillus iners,24),type Ⅳ (no Lactobacillus as the dominant bacteria,12),type Ⅴ (Lactobacillusjensenii,2).16S rDNA V1V2 region sequence analysis showed that of the 49 samples,13 belonged to type Ⅰ,type Ⅱ (1),type Ⅲ (23),type Ⅳ (8),type Ⅴ (2).Two methods of vaginal flora classification were consistent for 38 cases,consistent rate was 77.6％.Conclusion·Two methods analysis of vaginal flora showed the different results.If qPCR was used to classify the vaginal microbiome,it was necessary to consider the influence of relevant technical factors on the results.