二代基因测序技术在微生物鉴定中的应用

Application of Next-Generation Sequencing Technology in Microbial Identification

目的:运用二代基因测序技术建立微生物鉴定的方法。方法:对细菌16S rRNA基因的V1-V3区进行扩增,对真菌的ITS基因的ITS1、ITS2区进行扩增。扩增产物加标签后,用iSeq100进行测序,用配套的微生物鉴定系统进行数据分析,得出鉴定结果。结果:鉴定了22株环境菌和31株标准菌。在属水平上,鉴定结果与对照菌一致率为100%;在微生物种水平上鉴定结果与对照菌的一致率为81.1%。结论:运用二代基因测序技术成功建立了细菌、酵母菌、霉菌的鉴定方法,该方法鉴定成功率和准确性高、重现性好,可用于鉴定环境菌。

Objective:The method of microbial identification was established by next-generation gene sequencing technology.Methods:For bacterial,V1-V3 region of 16S rRNA gene was amplified.For fungal,ITS1 and ITS2 gene regions were amplified.After labeling,the amplified products were sequenced by iSeq100,and the data were analyzed by the microbial identification system to obtain the identification results.Results:22 strains environmental microorganisms and 31 strains standard microorganisms were identified.At the genus level,the concordance of identification results was 100%.At the species level,the concordance of identification results was 81.1%.Conclusion:The identification methods of bacteria,yeast and mold were successfully established by next-generation sequencing technology.The methods have high success rate,accuracy and reproducibility,can be used to identify environmental microorganisms.