用PCR技术鉴定犬传染性肝炎病毒强、弱毒株的研究  

Establishment of polymerase chain reaction for discrimination of virulent and attenuated infectious strains of canine hepatitis viruses

作　　者：王雷[1] 夏咸柱[1] 卫广森 邹啸环[1] 刘维全[1] 扈荣良[1] 黄耕[1] 

机构地区：[1]中国人民解放军军需大学,吉林长春130062 [2]辽宁益康生物制品厂,辽宁辽阳111000

出　　处：《畜牧与兽医》2002年第2期10-12,共3页Animal Husbandry & Veterinary Medicine

基　　金：全军医学科学研究"十五"计划重点课题 (0 1 Z 0 92 )

摘　　要：根据Genebank中发表的犬传染性肝炎病毒 (ICHV)标准强毒株Glaxo和人工驯化的弱毒疫苗株CLL的保守序列间大小不同 ,按照引物设计的原则 ,设计合成了一对通用引物。该对引物可由ICHV强毒株扩增出 569bp的片段 ,而弱毒株可扩增出 2 4 4bp的片段。对PCR产物分别进行电泳、酶切和测序 ,证明PCR产物片段大小、酶切位点和核苷酸序列与设计的产物完全一致。正常DK细胞上清和犬传染性喉气管炎病毒 (CAV 2 )强、弱毒株细胞培养物均不能被该引物扩增 ,说明具有良好的特异性。其检测到的病毒量分别为 1 5TCID50 、 31TCID50 、说明该技术具有很高的敏感性。可用于ICHV强。One pair of PCR primers were designed according to the sequences of the virulent strain Glaxo and attenuated strain CLL of canine adenovirus type I.The conserved regions of the viruient and the attenuated infectious canine hepatitis virus(ICHV)are different.After amplificating,a 569bp fragment was obtained from the virulent strain while 244bp fragment from the attenuated strain.The products of PCR were examined by electrophoresis,restriction enzyme digestion and sequencing assay.The results showed that the length of fragments,enzyme sites and sequence were as the same as those designed for Glaxo and CLL.No specific fragment was amplified from MDCK control cells and canine adenovirus 2 culture.The PCR of CAV?1 was proved to be sensitive and specific.It shows that this method may be used in discriminating the virulence strains of the ICHVs and clinical in diagnosis of ICHV infection.

关 键 词：PCR Ⅰ型犬腺病毒 毒力鉴定 技术鉴定 犬 传染性肝炎 病毒毒株 

分 类 号：S858.292[农业科学—临床兽医学]