猪链球菌长沙分离株的鉴定与16S rRNA系统进化分析  

Identification and 16SrRNA phylogenetic analysis of 2 Streptococcus suis strains isolated in Changsha City

作　　者：苏良[1] 欧新华[1] 杨柳青[1] 张如胜[1] 刘如春[1] 陈田木[1] 李亚曼[1] 孙边成[1] 

机构地区：[1]长沙市疾病预防控制中心,长沙410001

出　　处：《中国人兽共患病学报》2013年第6期547-550,共4页Chinese Journal of Zoonoses

基　　金：长沙市科技计划项目(No.K0902167-31);湖南省疾控中心微生物检验科对本研究的支持~~

摘　　要：目的对2株猪链球菌(Streptococcus suis,SS)长沙分离株进行分离培养和分子鉴定,并对其16SrRNA基因进行测序,阐明其系统进化关系。方法利用分离培养法对2株SS进行分离鉴定,同时对分离株16SrRNA基因进行PCR扩增和核苷酸序列测定,将测序结果提交GenBank,通过在线Blast同源性分析进行菌株鉴定,并与传统培养鉴定方法进行比较,利用Mega4.0软件构建SS长沙分离株系统进化树。结果成功扩增出2株菌的目的片段,通过测序后Blast同源性分析,证实2株菌株均为SS,且与传统鉴定方法结果一致,进化树显示长沙2株SS和国内外2型位于同一进化分支上,同四川的05ZYH33参考株亲缘关系最近。结论通过16SrRNA基因可以准确快速鉴定SS,可应用于应急检测,长沙2株猪链球菌属于SS2型,与其他SS2分离株16SrRNA基因同源性高,未发生变异。The aim of this study was to identify 2 strains of Streptococcus suis isolated in Changsha City and elucidate their phylogenetic relationship through the 16S rRNA. The 16S rRNA of 2 strains was amplified by polymerase chain reaction （PCR） through universal primer of 16S rRNA. The products of 16S rRNA of 2 strains were obtained successfully with length about 1 500 bp. The products were sequenced successfully then the sequences were submitted to GenBank. And the sequences were compared with related sequence through online BLAST in NCBI, showing the 2 strains of Streptococcus suis were Strep tococcus suis type 2. The identification result through the 16S rRNA was consistent with traditional method. Then the phyloge netic tree of 16S rRNA was constructed through the Mege 4.0 software. The phylogenetic tree showed that the 2 strains of Streptococcus suis were at the same evolutionary branch with most of Streptococcus suis type 2 and had the nearest phylogenetic relationship with Streptococcus suis 05ZYH33 in Sichuan Province. In conclusion, Streptococcus suis could be quickly and accu rately identified through the 16S rRNA and this method could be applied in rapid detection. The 2 strains of Streptococcus suis isolated in Chansha City were Streptococcus suis type 2 and was not variable in 16S rRNA gene.

关 键 词：猪链球菌 16S RRNA 鉴定 进化分析 

分 类 号：R378.12[医药卫生—病原生物学]