尼日利亚妇女健康和疾病中的阴道细菌组:16S rRNA宏基因组学研究

K. Anukam, N. Agbakoba, A. Okoli, C. Oguejiofor
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引用次数: 9

摘要

引言:关于什么细菌构成健康阴道和细菌性阴道病(BV)的争论仍未解决。黑人女性通常被安置在四级阴道群落中,因为缺乏以乳酸杆菌为主的微生物。我们试图使用16S rRNA宏基因组学方法来确定健康人和BV患者的阴道微生物群组成。材料和方法:28名女性提供阴道拭子进行Nugent评分。15人患有BV(Nugent评分7-10),13人正常(Nugent得分0-3)。在用MiSeq平台测序之前,提取DNA并使用定制的条形码引物扩增16S rRNA V4区域。将序列读数导入Illumina BaseSpace宏基因组学管道,用于16S rRNA识别。使用Greengenes数据库对不同分辨率级别的分类类别进行了分布。相似性聚类采用Manhattan主成分分析。结果:非BV受试者被12个分类门定殖,共代表182属357种。总的来说,在BV受试者中鉴定出23个门,代表388属和805种。在非BV受试者中,厚壁菌门占序列读数的95%,属以乳酸杆菌为主,种以脆乳杆菌为主,其次是变形菌门(3.78%)、放线菌门(0.74%)和细菌门(0.05%),变形杆菌属(1.48%)和Tenericutes属(1.25%)。结论:与已发表的报告相反,非BV健康的非洲黑人尼日利亚妇女的乳酸杆菌属是主要的微生物群。这项研究表明,BV受试者有不同比例的不同细菌,类似于世界其他地区的研究。
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Vaginal bacteriome of Nigerian women in health and disease: A study with 16S rRNA metagenomics
Introduction: The argument on what bacteria make up healthy vagina and bacterial vaginosis (BV) remain unresolved. Black women most often are placed in grade IV vaginal communities as lacking Lactobacillus-dominated microbes. We sought to determine the vaginal microbiota compositions of healthy and those with BV using 16S rRNA metagenomics methods. Materials and Methods: Twenty-eight women provided vaginal swabs for Nugent scoring. Fifteen had BV (Nugent score 7–10), whereas 13 were normal (Nugent score 0–3). DNA was extracted and 16S rRNA V4 region amplified using custom bar-coded primers prior to sequencing with MiSeq platform. Sequence reads were imported into Illumina BaseSpace Metagenomics pipeline for 16S rRNA recognition. Distribution of taxonomic categories at different levels of resolution was done using Greengenes databases. Manhattan principal component analysis was used for similarity clustering. Results: Non-BV subjects were colonized by 12 taxonomic phyla that represent 182 genera and 357 species. Overall, 23 phyla representing 388 genera and 805 species were identified in BV subjects. Firmicutes represented 95% of the sequence reads in non-BV subjects with Lactobacillus-dominated genera and Lactobacillus crispatus–dominated species, followed by Proteobacteria (3.78%), Actinobacteria (0.74%), and Bacteriodetes (0.05%). In BV subjects, Firmicutes represented 59% of the classified sequence reads, followed by Bacteroidetes (19%), Actinobacteria (15.8%), Fusobacteria (4.08%), Proteobacteria (1.48%), and Tenericutes (1.25%). Conclusion: Non-BV healthy Black African, Nigerian women had Lactobacillus genera as the predominant microbiota, contrary to published reports. The study shows that BV subjects had varying proportions of diverse bacteria similar to studies from other parts of the world.
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