多位点序列分型(MLST)在艾伯特埃希菌鉴定中的应用  

Application of multi-locus sequence typing(MLST)in the identification of Escherichia albertii

作　　者：刘祥[1] 许彦梅[2] 王斌[1] 邓建平[1] 肖波[1] 孙松松[1] 周阳[1] 熊衍文[2] 王红[1] 

机构地区：[1]四川省自贡市疾病预防控制中心,自贡643000 [2]中国疾病预防控制中心传染病预防控制所传染病预防控制国家重点实验室,北京102206

出　　处：《中国人兽共患病学报》2015年第11期1033-1036,共4页Chinese Journal of Zoonoses

基　　金：自贡市重点科技计划项目(No.2013S06);四川省卫生和计划生育委员会科研课题(No.150259)~~

摘　　要：目的探讨多位点序列分型技术在新病原艾伯特埃希菌发现与鉴定中的应用。方法采用MLST技术对生鲜肉中分离的30株疑似艾伯特埃希菌的7个管家基因进行PCR扩增、测序、DNAstar软件分析,核酸序列上传至大肠杆菌MLST数据库进行比对,确定其等位基因编号及基因序列型(ST型),并利用MEGA6.0软件Neighbor-joining法构建遗传进化树,与大肠埃希菌、志贺氏菌、艾伯特埃希菌和伤寒沙门氏菌参考菌株进行亲缘性分析。结果 30株菌可分为7种新序列型(16株)和4种已知序列型(14株),N-J分析显示与艾伯特埃希菌参考菌株具有高度亲缘性,而与大肠埃希菌、志贺氏菌、弗格森埃希菌和鼠伤寒沙门氏菌存在较大差异。结论 MLST在管家基因水平上准确地确定艾伯特埃希菌的种属关系,首次在国内发现艾伯特埃希菌的存在。We studied the application of multilocus sequence typing （MLST） in the discovery and identification of a new pathogen Escherichia albertii. MLST was performed according to the E. coli MLST database using seven housekeeping genes. The 7 housekeeping genes of 30 suspected Escherichia albertii strains isolated from retail raw meats were amplified, sequenced and analysed by DNA star software. The nucleotide sequences were uploaded to E. coli MLST database to confirm the allele number and the sequence type （ST type）. To analyse the phylogenetic relationships between the 30 strains and the reference strains of E. coli, E. fergusonii, E. albertii, Shigella spp. and Salmonella enterica serovar Typhi, the phylogenetic tree was constructed with the neighbor-joining method of MEGA 6. Results showed that 30 strains were classified into 7 new STs （16 strains） and 4 known STS （14 strains）, the analysis of N-J showed high homology with E. albertii strain, while were highly divergent from E. coli, Shigella sp. , E. fergusonii and Salmonella enterica serotype Typhi stains. MLST accurately determine the relationship between species of E. albertii at the level of housekeeping genes. And it is the first time of discove- ring and confirming the existence of E. albertii in China.

关 键 词：多位点序列分型技术(MLST) 艾伯特埃希菌 新病原 

分 类 号：R378[医药卫生—病原生物学]