摘要/Abstract
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
PDF全文下载地址:
点我下载PDF
