›› 2018, Vol. 38 ›› Issue (1): 71-.doi: 10.3969/j.issn.1674-8115.2018.01.013

• Original article (Clinical research) • Previous Articles     Next Articles

Comparison of real-time quantitative PCR and 16S rDNA sequencing for detection of female vaginal microbiome

ZHANG Ning1, 2, HAN Yang1, 2, LUO Fei3, ZHU Li-hong4, QIN Jin-hong2, JIANG Yan1   

  1. 1. Key Laboratory of Medical Microbiology and Parasitology, Guizhou Medical University, Guiyang 550025, China; 2. Department of Microbiology and Immunology, Shanghai Jiao Tong University College of Basic Medical Sciences, Shanghai 200025, China; 3. International Peace Maternity & Child Health Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200030, China; 4. Department of Gynecology, Huadong Hospital, Fudan University, Shanghai 200040, China
  • Online:2018-01-28 Published:2018-03-09
  • Supported by:
    Shanghai Jiao Tong University Biomedicine-Engineering Research Foundation, YG2016MS83; Natural Science Foundation of Shanghai, 17ZR1415900

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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 (Lactobacillus jensenii, 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.

Key words: vaginal flora, real-time quantitative PCR, 16S rDNA high throughput sequencing