青岛地区空肠弯曲菌的分离鉴定及Jej基因序列分析  

Isolation and Identification of Campylobacter jejuni Isolates in Qingdao and Analysis of Jej Gene Sequences

作　　者：张维[1,2] 盖文燕[3] 王娟[3] 王君玮[3] 范红结[1] 曲志娜[3] 黄秀梅[3] 翟海华[3] 

机构地区：[1]南京农业大学动物医学院,江苏南京210095 [2]青岛易邦生物工程有限公司,山东青岛266114 [3]中国动物卫生与流行病学中心,山东青岛266032

出　　处：《河南农业科学》2014年第7期138-143,168,共7页Journal of Henan Agricultural Sciences

基　　金：农业部"引进国际先进农业科学技术"重点项目-948项目[2011-G14(2)]

摘　　要：为调查青岛地区空肠弯曲菌（C.jejuni）的流行状况,采集青岛地区不同分离地点的鸡盲肠棉拭子及内容物98份,经分离鉴定得到23株C.jejuni,分离率达到23.47%。应用C.jejuni的马尿酸酶（Jej）基因作为分子标记,对分离到的23株C.jejuni分离株和4株C.jejuni上海分离株进行分析,并与8株已知的C.jejuni序列进行相似性比较,构建系统进化树,对其系统进化关系进行分析。结果表明,分离菌株及已知菌株均能扩增出774bp的Jej基因片段,得到11个不同的序列,有11个核苷酸变异位点,变异率为0.13%～1.42%;基于Jej基因构建的进化树表明,分离株分为2个分支,其中4株分离株与C.J-NCTC11168和C.J-IA3902亲缘关系近,1株与C.JRM1221、C.J-S3亲缘关系近,2株与上海分离株亲缘关系比较近;2株与C.J-81116、C.J-M1组成另外一个分支。To research the prevalence of C.jejuni in Qingdao for the different geographical distribution genetic variation study,23 strains of C.jejuni were isolated from 98 chicken appendix cotton slips and contents,the isolation rate was 23.47%.The C.jejuni hippuricase（Jej）gene was used as a molecular marker to identify 23 isolates from Qingdao area and four Shanghai isolates.The Jej gene fragment was cloned and sequenced fromC.jejuni samples.Sequence analysis,similarity comparison with the known eight C.jejuni and phylogenetic analysis were performed to study the source and phylogenetic relationship.Results showed that 774 bp of the Jej gene fragment were successfully amplified from all of the 27 isolates.Sequence analysis indicated that the sequences could be divided into eleven types with a total of eleven nucleotide variant sites（variant rate from 0.13% to 1.42%）.The phylogenetic tree based on Jej sequence showed that all Qingdao isolates could be divided into two branches.One branch was that four Qingdao isolates showed the closest relationship with C.J-NCTC11168 and C.J-IA3902,C.J-RM1221,C.J-S3 and one Qingdao isolate showed the closest relationship,followed by two Qingdao isolates and Shanghai isolates as the closest relationship.Two Qingdao isolates,C.J-81116 and C.J-M1 constituted another branch.

关 键 词：青岛 空肠弯曲菌 马尿酸酶基因 序列分析 系统进化 溯源 

分 类 号：S855.1[农业科学—临床兽医学]