牛荚膜血清A型巴氏杆菌病的分子流行病学调查  

Molecular epidemiological survey of capsular serotype A Pasteurellosis in cattle

作　　者：马文戈[1] 姜志刚[1] 于力[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/大动物病研究室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2010年第5期360-364,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家科技支撑计划(2006BAD04A09);黑龙江省科技攻关重大项目专题(GA06B202)

摘　　要：本研究针对多杀性巴氏杆菌(Pm)特异性基因kmt1,进化保守基因16S rRNA基因,以及荚膜血清型A、B、D、E和F型对应的荚膜生物合成基因hayD-hayC、bcbD、dcbF、ecbJ和fcbD,设计特异性PCR扩增引物,采用PCR方法扩增相应基因和进行序列测定,并对分离菌株进行分子鉴定和同源性分析。结果显示,来自不同地区的分离菌株均含有Pm种特异性基因kmt1、A型荚膜生物合成基因hayD-hayC和16S rRNA基因。不同地区的分离株,kmt1基因的同源性为100%;A型荚膜生物合成基因hayD-hayC同源性大于99.9%;与国外牛源分离株的hayD-hayC基因的同源性大于98%;不同地区分离株的16S rRNA基因的同源性为100%,而与英国牛源分离株Pm338的16S rRNA基因的同源性高达99.93%。这些结果表明,在我国6个省市流行的牛出血性败血症由同一来源的荚膜血清A型多杀性巴氏杆菌所致,与英国牛源A型分离株Pm338具有共同的进化来源。Pasteurella multocida（Pm） is a heterogeneous causative species in domesticated and wild animals.Current capsular serotyping methods classify Pm into five groups A,B,D,E and F based on multiplex PCR assay.Serotype A,B and E strains infect cattle causing septicemia or respiratory disease.In this study,Pm isolates of serotype A isolated from pneumonic cattles in Heilongjiang Tianjin,Jilin,Neimenggu,Ningxia and Shandong provinces were subjected to PCR using primers specific to the species-specific gene kmt1,capsule biosynthesis genes hayD-hayC（A serotype）,bcbD（B serotype）,dcbF（D serotype）,ecbJ（E serotype）,fcbD（F serotype） and 16S rRNA gene.The PCR products of relevant genes were sequenced and compared.The results showed that the kmt1 genes and 16S rRNA genes of these islates were 100 % identical,and the capsule biosynthesis genes hayD-hayC（serotype A） derived from different regional isolates shared more than 99.9 % identities.The 16S rRNA gene of those Pm isolates had higher sequence identities with England bovine isolate Pm338 than that of other isolates.Phylogenetic analysis of 16S rRNA genes revealed that the all Pm isolates derived from different regions of China originated from a common ancestor together with the England isolate Pm338.

关 键 词：多杀性巴氏杆菌分离株 荚膜血清A型 16S RRNA kmt1基因 进化分析 

分 类 号：S852.61[农业科学—基础兽医学]