Department of Obstetrics and Gynecology, Kansai Medical University Graduate School of Medicine, Osaka, Japan.
HORAC Grand Front Osaka Clinic, Osaka, Japan.
BMC Med Genomics. 2022 Mar 25;15(1):68. doi: 10.1186/s12920-022-01218-8.
It has been suggested that the local microbiota in the reproductive organs is relevant to women's health and may also affect pregnancy outcomes. Analysis of partial 16S ribosomal RNA (rRNA) gene sequences generated by short-read sequencers has been used to identify vaginal and endometrial microbiota, but it requires a long time to obtain the results, making it unsuitable for rapid bacterial identification from a small specimen amount in a clinical context.
We developed a simple workflow using the nanopore sequencer MinION that allows high-resolution and rapid differentiation of vaginal microbiota. Vaginal samples collected from 18 participants were subjected to DNA extraction and full-length 16S rRNA gene sequencing with MinION.
The principal coordinate analysis showed no differences in the bacterial compositions regardless of the sample collection method. The analysis of vaginal microbiota could be completed with a total analysis time of approximately four hours, allowing same-day results. Taxonomic profiling by MinION sequencing revealed relatively low diversity of the vaginal bacterial community, identifying the prevailing Lactobacillus species and several causative agents of bacterial vaginosis.
Full-length 16S rRNA gene sequencing analysis with MinION provides a rapid means for identifying vaginal bacteria with higher resolution. Species-level profiling of human vaginal microbiota by MinION sequencing can allow the analysis of associations with conditions such as genital infections, endometritis, and threatened miscarriage.
有人认为生殖器官中的局部微生物群与女性健康有关，也可能影响妊娠结局。通过短读长测序器生成的部分 16S 核糖体 RNA（rRNA）基因序列分析已被用于鉴定阴道和子宫内膜微生物群，但需要很长时间才能获得结果，因此不适合在临床环境中从少量标本中快速鉴定细菌。
我们开发了一种使用 MinION 纳米孔测序器的简单工作流程，该流程允许对阴道微生物群进行高分辨率和快速区分。对 18 名参与者采集的阴道样本进行 DNA 提取和全长 16S rRNA 基因 MinION 测序。
主坐标分析表明，无论采用何种样本采集方法，细菌组成均无差异。阴道微生物群的分析可以在大约四个小时的总分析时间内完成，从而实现当天出结果。通过 MinION 测序进行的分类分析显示，阴道细菌群落的多样性相对较低，鉴定出了主要的乳酸杆菌属物种和几种细菌性阴道病的病原体。
使用 MinION 进行全长 16S rRNA 基因测序分析提供了一种快速识别阴道细菌的方法，具有更高的分辨率。通过 MinION 测序对人阴道微生物群进行种属水平分析，可以分析与生殖器感染、子宫内膜炎和先兆流产等疾病的关联。
