上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2018, Vol. 38 ›› Issue (1): 71-.doi: 10.3969/j.issn.1674-8115.2018.01.013

• 论著（临床研究） • 上一篇    下一篇

运用实时定量PCR 和16S rDNA 测序法分析女性阴道菌群的结果比较

张宁1, 2，韩阳1, 2，骆菲3，朱丽红4，秦金红2，江滟1   

  1. 1. 贵州医科大学贵州省普通高校病原生物学特色重点实验室，贵阳550025；2. 上海交通大学 基础医学院免疫学与微生物学系，上海200025；
    3. 上海交通大学 医学院附属国际和平妇幼保健院，上海 200030；4. 复旦大学附属华东医院妇科，上海200040
  • 出版日期:2018-01-28 发布日期:2018-03-09
  • 通讯作者: 江 滟，电子信箱：624057523@qq.com。
  • 作者简介:张 宁（1985—），女，硕士生；电子信箱：zhangn_gy@163.com。
  • 基金资助:
    上海交通大学医工交叉研究基金（YG2016MS83）；上海市自然科学基金（17ZR1415900）

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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摘要： 目的· 比较实时定量PCR（qPCR）法与16S rDNA 测序法分析女性阴道菌群类型的结果。方法· 选取45 岁以下育龄期健康
女性49 名，采集阴道上1/3 处分泌物，经提取阴道标本全基因组DNA 后，分别运用16S rDNA V1V2 域测序和基于22 种常见阴道
细菌设计的引物进行qPCR 分析各样本菌群类型，比较2 种检测结果的异同。结果· 参考依据优势菌类型的阴道菌群分类标准，基于
qPCR 方法分析结果为Ⅰ型（卷曲乳杆菌为优势菌）9 例，Ⅲ型（惰性乳杆菌为优势菌）24 例，Ⅳ型（无乳杆菌为优势菌）12 例，Ⅴ
型（詹氏乳杆菌为优势菌）为2 例。基于16S rDNA V1V2 域进行测序分析的结果为Ⅰ型13 例，Ⅱ型（格氏乳杆菌为优势菌）1 例，
Ⅲ型23 例，Ⅳ型8 例，Ⅴ型2 例。2 种方法对阴道菌群分型一致的为38 例，一致率为77.6%。结论· 2 种方法分析得到的阴道菌群分
类结果有差异；若采用qPCR 对阴道菌群进行分类，还需考虑相关的技术因素对结果的影响。

关键词: 阴道菌群, 实时定量PCR, 16S rDNA 高通量测序

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