不同16S rDNA高变区选择对细菌性阴道病菌群研究差异比较  

Comparison of different 16S rDNA hypervariable regions selection for the microbiome study of bacterial vaginosis

作　　者：刘瑛[1,2] 张琼琼 张蕾[1,2] 王颖[1,2] 吕涛[1,2] 陶址 黄振宇[1,2] 廖秦平[1,2] Liu Ying;Zhang Qiongqiong;Zhang Lei(Department of Obstetrics and Gynecology,Beijing Tsinghua Changgung Hospital,School of Clinical Medicine,Tsinghua University,Beijing 102218;School of Clinical Medicine,Tsinghua University,Beijing 100084)

机构地区：[1]清华大学附属北京清华长庚医院妇产科,北京102218 [2]清华大学临床医学院,北京100084

出　　处：《现代妇产科进展》2019年第11期804-807,共4页Progress in Obstetrics and Gynecology

基　　金：国家自然科学基金(No:81671409);北京市医院管理局临床医学发展专项经费(No:XMLX201605)

摘　　要：目的:分析不同16S rDNA高变区对细菌性阴道病菌群研究的差异,为选择合适的高变区扩增细菌性阴道病菌群提供理论依据。方法:选取2017年10月至2018年5月于北京清华长庚医院妇科门诊就诊的28例健康女性和10例细菌性阴道病患者,收集患者的阴道分泌物,提取细菌总DNA,分别用两对引物27F/338R和341F/806R对细菌16S rDNA的V1/V2区和V3/V4区进行PCR扩增后测序,通过分析两个高变区扩增后Alpha和Beta多样性及菌群结构的差异,比较两个高变区在细菌性阴道病菌群研究上的优势。结果:16S rDNA的V3/V4区扩增的引物较V1/V2区能扩增出更多的阴道菌种。V1/V2区扩增BV样本的加德纳菌丰度偏低。通过V3/V4区的扩增,无法将阴道健康样本中卷曲乳杆菌与母鸡乳杆菌区分开。结论:16S rDNA的V3/V4区较V1/V2区更适用于细菌性阴道病菌群结构及丰度分析。Objective:To analyze the microbiome difference of bacteria vaginosis in different 16s rDNA hypervariable regions,to provide theoretical basis for selecting suitable hypervariable regions to amplify the microbiome of bacteria vaginosis.Methods:Vaginal secretions were collected from 28 healthy women and 10 bacterial vaginosis women,who were admitted to the gynecological clinic of Beijing Tsinghua Changgung Hospital from Oct.2017 to May 2018.After extracting the total bacterial DNA,primer 27F/338R and primer 341F/806R were used to sequence the V1/V2 region and V3/V4 region of 16S rDNA by PCR amplification.Compared the advantages of two hypervariable regions in the microbiome study of bacteria vaginosis by analyzing the diversity of Alpha and Beta and the difference of bacterial structure.Result:Primers for V3/V4 region of 16S rDNA amplified more vaginal strains than V1/V2 region.The abundance of Gardnerella in BV samples amplified by V1/V2 region was low.Amplification by the V3/V4 region could not distinguish Lactobacillus crispatus and Lactobacillus gallinarum in healthy vaginal samples.Conclusions:The V3/V4 region of 16s rDNA is more suitable for the analysis of the structure and abundance of bacterial vaginosis than V1/V2 region.

关 键 词：16S rDNA高通量测序 引物 阴道菌群 菌群多样性 

分 类 号：R711.3[医药卫生—妇产科学]