纳米孔测序技术检测正常人群咽拭子微生物本底的初步分析

Preliminary analysis of microbiological background in normal human throat swab using nanopore sequencing technology

目的分析咽拭子中存在多种少量的机会致病性微生物(即病原体本底),为测序数据提供初步分析结果。方法利用牛津纳米孔测序技术(oxford nanopore technologies)对12个健康人的咽拭子样本进行测序并分析微生物序列。结果咽拭子样本中主要是常见的宿主共生菌群,具体为拟杆菌门(29.000%~55.000%)、变形菌门(12.000%~24.000%)、厚壁菌门(4.000%~7.000%)、梭杆菌门(2.000%~3.000%)和放线菌门(0.300%-1.000%)等。在拟杆菌门中,普雷沃菌(82.000%~93.000%)、黄杆菌(2.000%~9.000%);变形菌门中,γ-变形菌(61.000%~79.000%)、β-变形菌(5.000%~28.000%);厚壁菌门中,链球菌(35.000%~61.000%)、葡萄球菌(2.000%~7.000%)、韦荣球菌(17.000%~33.000%)、梭菌(2.000%~5.000%)、微单胞菌(0.6000%~1.000%);梭杆菌门中,梭杆菌(54.000%~84.000%)、纤毛菌(9.000%~25.000%);放线菌门中,放线菌(19.000%~47.000%)。最后,还在样本中检测出了少量的病毒,鉴定结果显示这些病毒主要是寄生在口腔中的细菌噬菌体。结论研究为临床宏基因组测序技术在呼吸道样本中病原体分析奠定了基础。

Objective This research aimed to reveal the abundance of opportunistic pathogenic microorganisms in throat swabs(i.e.pathogen background),and conduct a preliminary analysis of sequencing data.Methods Throat swab samples obtained from 12 healthy people were sequenced using oxford nanopore technologies,and the sequencing data was analyzed to establish a pathogen background database.Results Microorganisms detected in throat swab samples were mainly composed of host commensal flora,including Bacteroides(29.000%-55.000%),Proteobacteria(12.000%-24.000%),Firmicutes(4.000%-7.000%),Fusobacteria(2.000%-3.000%),Actinobacteria(0.300%-1.000%)and so on.Among Bacteroidetes,Prevotella and Flavobacterium were detected accounting for 82.000%~93.000%,2.000%~9.000%,respectively.whileγ-proteobacteria(61.000%-79.000%)andβ-proteobacteria(5.000%-28.000%)were detected belonging to Proteobacteria,and among Firmicutes,Streptococcus(35.000%-61.000%),Staphylococcus(2.000%-7.000%),Veillonella(17.000%-33.000%),Clostridium(2.000%-5.000%),Micromonas(0.600%-1.000%)were identified;while for Fusobacteria,Fusobacterium(54.000%-84.000%),Ciliates(9.000%-25.000%),and finally,among those belonged to Actinobacteria,actinomycetes accounting for 19.000%-47.000%.In addition,a small number of viruses were also detected in the throat swab samples,the identification results showed these viruses were mainly bacteriophages parasitizing in the oral cavity.Conclusion This study systematically analyzed the pathogen background in normal throat swabs and provided a valuable foundation for the analysis of clinical metagenomic sequencing technology in respiratory samples.