Diao Yutao, Fang Xueqiang, Xia Qing, Chen Shouqiang, Li Hao, Yang Yanfang, Wang Yan, Li Huiqing, Cui Jia, Sun Xiaomeng, Zhao Zhongtang
Department of Epidemiology and Health Statistics, School of Public Health, Shandong University, China.
J Obstet Gynaecol Res. 2011 Oct;37(10):1438-46. doi: 10.1111/j.1447-0756.2011.01564.x. Epub 2011 Jun 16.
The aim of this study was to characterize the different structures of microbial communities between 20 healthy women and 17 bacterial vaginosis (BV)-positive women of reproductive age using denaturing gradient gel electrophoresis (DGGE).
Vaginal samples from 17 BV-positive and 20 BV-negative women were subjected to DNA extraction, and amplified with eubacterial 16S rRNA gene-specific primers via polymerase chain reaction. The polymerase chain reaction products were separated using DGGE. Bands were excised, re-amplified, purified and sequenced. DNA sequences were compared with GenBank database. Phylip software packages were used to calculate sequencing data and form a phylogenetic tree to identify the genetic relations for microbiota inhabited in vaginal ecosystems of BV-positive women.
In total, 28 kinds of organisms were detected that comprised BV(+) vagina microbial community, varying from three to nine kinds with an average of 5.71 kinds per woman. Only seven species were detected in BV(-) women, ranging between one and five species with an average of 2.40 species per woman, which was significantly lower than that detected in BV(+) women (t = 7.39, P < 0.001). A strain of Uncultured Lactobacillus sp. clone EHFS1_S05c (29/37; 78.38%) was most commonly presented in both BV-negative and BV-positive women, but the mean proportion of this Lactobacillus sp. strain to the whole microbial population colonized in the vaginal tract of BV(-) women was sharply higher than that calculated from BV(+) women (t = 2.92, P < 0.01).
The findings indicate further diversity in the category of vaginal microorganisms associated with BV. The presence of Gardnerella vaginalis is not necessary as a sign for gynecologists to determine whether or not a woman is affected by BV.
本研究旨在通过变性梯度凝胶电泳(DGGE)对20名健康育龄女性和17名细菌性阴道病(BV)阳性育龄女性的微生物群落不同结构进行表征。
对17名BV阳性和20名BV阴性女性的阴道样本进行DNA提取,并通过聚合酶链反应使用真细菌16S rRNA基因特异性引物进行扩增。聚合酶链反应产物通过DGGE进行分离。切下条带,重新扩增、纯化并测序。将DNA序列与GenBank数据库进行比较。使用Phylip软件包计算测序数据并形成系统发育树,以确定BV阳性女性阴道生态系统中栖息的微生物群的遗传关系。
共检测到28种构成BV(+)阴道微生物群落的生物体,每位女性的种类从3种到9种不等,平均每位女性5.71种。在BV(-)女性中仅检测到7种,范围在1种到5种之间,平均每位女性2.40种,明显低于BV(+)女性中检测到的种类(t = 7.39,P < 0.001)。一株未培养的乳酸杆菌属克隆EHFS1_S05c(29/37;78.38%)在BV阴性和BV阳性女性中均最常见,但该乳酸杆菌属菌株在BV(-)女性阴道中定殖的整个微生物群体中的平均比例明显高于BV(+)女性计算得出的比例(t = 2.92,P < 0.01)。
研究结果表明与BV相关的阴道微生物类别存在进一步的多样性。阴道加德纳菌的存在并非妇科医生确定女性是否患有BV的必要标志。