基于高通量测序的粉尘螨体内细菌多样性研究

Bacterial community diversity in Dermatophagoides farinae using high-throughput sequencing

目的探讨粉尘螨(Dermatophagoides farinae)体内细菌多样性。方法取实验室人工饲养的粉尘螨,采用Illumina PE250高通量测序平台扩增测序细菌16S核糖体RNA(16S ribosomal RNA,16S rRNA)基因V4区,获得的原始序列经质控过滤后得到有效序列。采用Usearch软件、Silva数据库、Mothur软件对有效序列进行操作分类单元(operational taxonomic units,OTU)聚类并分析细菌群落组成与alpha多样性指数。结果共获得187616条有效序列,根据序列相似性>97%进行聚类,得到469个OTUs。OTU注释结果表明,粉尘螨体内细菌分属于26个门、43个纲、100个目、167个科、284个属。在门分类阶元水平,粉尘螨体内细菌主要注释到变形菌门、厚壁菌门、拟杆菌门、放线菌门和酸杆菌门,其中变形菌门为优势菌门;在属分类阶元水平,罗尔斯通氏菌属、立克次体目未确定属、葡萄球菌属和鞘氨醇单胞菌属为优势菌属。在非优势菌属中,发现沃尔巴克氏体。结论粉尘螨体内细菌群落组成具有较高多样性,个体间细菌群落多样性与丰度存在差异。

Objective To investigate the bacterial community diversity in Dermatophagoides farinae.Methods Laboratorycultured D.farinae was collected,and the composition of microbial communities was determined by sequence analyses of the V4region in the bacterial 16S ribosomal RNA(16S r RNA)gene on an Illumina PE250 high-throughput sequencing platform.Following quality control and filtering of the raw sequence files,valid reads were obtained and subjected to operational taxonomic units(OTU)clustering and analysis of the composition of microbial communities and alpha diversity index using the Usearch software,Silva database,and Mothur software.Results A total of 187616 valid reads were obtained,and 469 OTUs were clustered based on a sequence similarity of more than 97%.OTU annotation showed that the bacteria in D.farinae belonged to 26 phyla,43 classes,100 orders,167 families and 284 genera.The bacteria in D.farinae were mainly annotated to five phyla of Proteobacteria,Firmicutes,Bacteroidota,Actinobacteriota,and Acidobacteriota,with Proteobacteria as the dominant phylum,and mainly annotated to five dominant genera of Ralstonia,norank-f-Mitochondria,Staphylococcus and Sphingomonas,with Wolbachia identified in the non-dominant genus.Conclusions A high diversity is identified in the composition of the bacterial community in D.farinae,and there are differences in bacterial community diversity and abundance among D.farinae.