健康妇女及3例细菌性阴道病患者阴道菌群的非培养分析

Culture-independent Analysis of Vaginal Microbial Communities in Healthy Women and Three Patients with Bacterial Vaginosis

摘要:

根据Amsel标准及Nugent标准, 确诊筛选健康妇女及细菌性阴道病(bacterial vaginosis, BV)患者各3例。提取其阴道分泌物样本的总DNA, 构建16S rRNA基因克隆文库, 并对阳性克隆进行ARDRA和测序分析。结果表明, 健康妇女样本的基因文库中, 分别以卷曲乳酸杆菌(L. crispatus)和惰性乳酸杆菌(L. iners)的克隆子占较大比例, 另外存在少量的阴道乳酸杆菌(L. vaginalis)或詹氏乳酸杆菌(L. jensenii)的克隆子。BV患者样本的基因文库中, 克隆子所代表的菌种类型明显增多, 但均以阴道加德纳氏菌(Gardnerella vaginalis)和阴道阿托波氏菌(Atopobium vaginae)的克隆子占较大比例, 且无乳酸杆菌克隆子。说明健康妇女阴道菌群的种类单一, 以乳酸杆菌占优势, L. iners为优势菌种之一; BV患者阴道菌群的种类复杂多样, 但均以Gardnerella vaginalis及Atopobium vaginae共同占优势。

关键词:细菌性阴道病, 16S rRNA基因, 克隆文库, 菌群

Abstract:

Three healthy women and three patients with BV were selected and diagonsed respectively according to the Amsel and Nugent criteria. Total microbial community DNA was extracted from their vaginal discharges, and 16S rRNA gene clone libraries were constructed. ARDAR fingerprinting patterns and sequences analysis of positive clones from each library showed that clones representing Lactobacillus crispatus and L. iners accounted for a major proportion respectively in the libraries for healthy women, where L. vaginalis and L. jensenii clones were present in a small amount. In the libraries for patients with BV, the microbial varieties increased remarkably, but both Gardnerella vaginalis and Atopobium vaginae clones constituted a greater portion, while Lactobacillus clones were not detected. It is therefore concluded that the vaginal microflora of healthy women is simple, predominated by single Lactbacillus species, and L. iners is one of the dominant species. The vaginal microflora of BV patients is complicated and various, but predominated by both Gardnerella vaginalis and Atopobium vaginae.

Key words:Bacterial vaginosis, 16S rRNA gene, Clone library, Community