犬瘟热病毒野毒株与疫苗株联合RT-PCR鉴别方法的建立  

Establishment of a multiplex RT-PCR assay for differentiation of field canine distemper virus from vaccine strains

作　　者：易立[1] 程世鹏[1] 闫喜军[1] 王建科[1] 罗彬[1] 赵建军[1] 张海玲[1] 吴威[1] 

机构地区：[1]中国农业科学院特产研究所,吉林吉林132109

出　　处：《中国兽医科学》2009年第7期617-621,共5页Chinese Veterinary Science

基　　金：吉林省科技发展计划项目(20080213)

摘　　要：为了建立快速、有效的鉴别犬瘟热病毒(CDV)野毒株与疫苗株的诊断方法,根据CDV野毒株与疫苗株N基因的保守区域以及H基因的多变区域设计特异性引物,建立了区分CDV野毒株与疫苗株的联合RT-PCR方法。结果表明,针对N基因的RT-PCR方法能同时检测CDV野毒株与疫苗株;而针对H基因的RT-PCR方法只能检测CDV野毒株,不能检测CDV疫苗株。根据2次RT-PCR反应结果,能够很好地区分CDV野毒株与疫苗株。同时,经过病毒电镜观察和H基因扩增片段的克隆测序结果的比对,进一步确认了该方法的特异性。表明,该方法具有快速、特异、实用等优点,可作为CDV鉴别诊断的有效手段。In order to establish a simple and rapid assay for differentiating canine distemper virus （CDV） field strains from vaccine strains,a multiplex RT-PCR assay was developed by setting up primers for N and H genes of CDV. The RT-PCR based on N gene could detect all CDV strains, and the RT-PCR based on H gene could only detect CDV field strains. Using the two RT-PCR,CDV field strains were differentiated easily from vaccine strains. Finally,the observation of electron microscope and the sequencing of H gene fragment amplified by RT-PCR confirmed further the specificity of the two RT-PCR assays. The results showed that the developed multiplex RT-PCR could be used for quick, specific and useful differentiation of CDV field strains from vaccine strains.

关 键 词：犬瘟热病毒 反转录聚合酶链式反应 野毒株 疫苗株 鉴别诊断 

分 类 号：S852.659.5[农业科学—基础兽医学] Q503[农业科学—兽医学]