2007-2020年上海市布鲁氏菌分离株分型方法研究  

Molecular characteristics of Brucella isolates in Shanghai,2007-2020

作　　者：高芬 陈洪友[1] 屠丽红[1] 庄源[1] 张曦[1] 陈敏[1] Gao Fen;Chen Hongyou;Tu Lihong;Zhuang Yuan;Zhang Xi;Chen Min(Shanghai Municipal Center for Disease Control and Prevention,Shanghai 200336,China)

机构地区：[1]上海市疾病预防控制中心,上海200336

出　　处：《疾病监测》2021年第7期696-701,共6页Disease Surveillance

基　　金：上海市卫生健康委员会科研课题(No.20194Y0397);国家科技重大专项(No.2018ZX10305409–003–007);上海市公共卫生优秀人才培养计划(No.GWV-10.2-YQ07)。

摘　　要：目的对2007—2020年上海市人间布鲁氏菌分离株进行分子特征分析,了解菌株基因分型和聚类,为布鲁氏菌病(布病)的预防和控制提供依据。方法收集2007—2020年上海市人感染布鲁氏菌临床分离株20株;采用生化方法和AMOS-PCR进行鉴定;运用多位点序列分型(MLST)、多位点可变数目串联重复序列分析(MLVA)和基于全基因组测序的单核苷酸多态性(SNP)分析3种方法进行分子分型;并与参考株进行聚类分析。结果20株布鲁氏菌均为ST8型羊种布鲁氏菌;被MLVA-8分为4个基因型,MLVA-16分为17个基因型,panel 2B具有多态性;SNP分析表明,20株临床分离株亲缘关系较近,与羊种布鲁氏菌聚类。结论2007—2020年上海市人感染布病分离株主要为ST8型羊种布鲁氏菌,分子分型方法可为上海市布鲁氏菌的溯源研究提供依据。Objective To investigate the molecular characteristics of Brucella isolates in Shanghai from 2007 to 2020,understand the genotyping and clustering of the strains,and provide evidence for the prevention and treatment of brucellosis.Methods A total of 20 clinical Brucella strains were isolated in Shanghai from 2007 to 2020.The strains were preliminarily identified by the biochemical method and AMOS-PCR.The multilocus sequence typing(MLST),multilocus variable number tandem repeat analysis(MLVA)and whole genome sequencing(WGS)based single nucleotide polymorphism(SNP)analysis were used to for molecular typing,and cluster analysis was performed with reference strains.Results By MLST,the 20 isolates were identified as Brucella melitensis,belonging to sequence types(ST)8.They were typed into 4 genotypes by MLVA-8 and 17 genotypes by MLVA-16,while locus of panel 2B showed polymorphism.WGS-based SNP analysis demonstrated that all the clinical strains were in cluster of B.melitensis,and some of which shared close phylogenetic relationship.Conclusion The strains isolated from human brucellosis cases in Shanghai from 2007 to 2020 were mainly ST8 B.melitensis.Molecular typing can provide evidence for tracing of human brucellosis in Shanghai.

关 键 词：布鲁氏菌属 多位点序列分析 多位点可变数目串联重复序列分析 全基因组测序 基因型 

分 类 号：R211[医药卫生—中医学] R378.5