奶牛传染性鼻气管炎病毒gG基因PCR检测方法的建立  

Establishment of PCR Method to Detect gG Gene of Infectious Bovine Rhinotracheitis Virus

作　　者：王冰[1,2,3] 张敏敏[1,3] 邹新峰[1,3] 郭爱珍[1,3] 

机构地区：[1]华中农业大学农业微生物学国家重点实验室,湖北武汉430070 [2]华中农业大学动物科技学院,湖北武汉430070 [3]华中农业大学动物医学院,湖北武汉430070

出　　处：《中国畜牧兽医》2011年第10期94-98,共5页China Animal Husbandry & Veterinary Medicine

基　　金：农业公益性行业科研专项(200803018;201003060);现代农业(肉牛)产业技术体系专项经费资助

摘　　要：本试验旨在建立一种PCR技术,既能快速检测牛传染性鼻气管炎病毒,又能区分同属病毒牛疱疹病毒5型和伪狂犬病病毒。根据基因库中牛传染性鼻气管炎病毒gG基因的特异性引物,建立PCR方法,对牛传染性鼻气管炎病毒参考毒株和阳性样本进行扩增,结果均能扩增出一条463bp的特异性条带;对同属的牛疱疹病毒5型进行扩增,获得651bp和431bp两条带;对同属伪狂犬病病毒进行扩增,获得493bp的条带;而非相关病毒(如猪呼吸与繁殖综合征病毒等),不能扩增出条带。对牛传染性鼻气管炎病毒的检测灵敏度为2×10-3 PFU/mL。鉴于该方法具有良好的灵敏度和特异性,将在牛疱疹病毒感染诊断和标记疫苗免疫后的鉴别诊断方面具有良好的应用前景。The specific primers were designed according to the published sequence of the infectious bovine rhinotracheitis virus（IBRV）gG gene.The method could amplify a specific IBRV fragment of 463 bp for IBRV reference strain and the IBRV positive samples.For bovine herpesvirus 5,it amplified two bands of 651 bp and 431 bp;for pseudorabies,the PCR product is a fragment of 493 bp.However,this PCR did not yield any product for non-related viruses such as porcine reproductive and respiratory syndrome virus.The sensitivity for IBRV was 2×10-3 PFU/mL.Since this PCR has good sensitivity and specificity,it would have a wide application in diagnosis of bovine herpesvirus infection and differential detection among the members of herpesviruses and between natural infection and vaccination of gG deleted marker vaccine.A PCR method was established successfully to detect infectious bovine rhinotracheitis virus（IBRV） and differentiate bovine herpesviruses.

关 键 词：牛传染性鼻气管炎病毒 PCR 牛疱疹病毒 gG基因 

分 类 号：Q78[生物学—分子生物学]