副猪嗜血杆菌的分离鉴定及其OmpP5基因序列分析

摘要

In order to study the genetic variation of the OmpP5 gene of Haemophilus parasuis in Liaoning province,this study was carried out from the pig's lungs in Fuxin,Liaoning province,which was suspected of suffering from Haemophilus parasuis.Gram staining,biochemical identification,satellite phenomenon and Hps 1 6 S rRNA PCR detection identified as Haemophilus parasuis.The OmpP5 gene was amplified, cloned and sequenced by the design of a pair of specific primers based on the published Hps OmpP5 gene. The OmpP5 gene sequence with length of 1 1 1 6 bp was obtained.The sequence was compared with the OmpP5 gene of the published Hps reference strain for homology comparison and phylogenetic tree analy-sis.The homology of the nucleotide was 89.8%-99.5%,and the isolate was the same as the reference strain№4,which indicated that OmpP5 had some conservatism,and the isolate was close to №4.The results pro-vided a reference for the study and pathogenesis of Hps virulence genes in Liaoning province.%为研究辽宁省副猪嗜血杆菌(Hps)OmpP5基因的遗传变异规律,从辽宁省阜新市某猪场疑似患有副猪嗜血杆菌病的患猪肺部采取组织病料进行细菌分离,通过革兰染色、生化鉴定、卫星现象及 Hps 16 S rRNA PCR检测,鉴定为副猪嗜血杆菌.根据已发表的 Hps OmpP5基因设计1对特异性引物,对分离株OmpP5基因进行扩增、克隆及测序,得到长度为1116 bp的 OmpP5基因序列.将该序列与已发表 Hps参考株的 OmpP5基因进行同源性比对及系统发育树分析,发现核苷酸同源性为89.8%~99.5%,分离株与参考菌株№4在同一分支,表明 OmpP5基因具有一定的保守性,分离株和№4亲缘关系较近.研究结果为阐明辽宁省 Hps毒力基因及致病机理提供了参考依据.