犬瘟热病毒和犬腺病毒2型双重PCR方法的建立及应用  

Establishment and rudimentary application of a duplex-PCR for Detection of CAV-2-CDV

作　　者：耿庆华[1] 林颖[1] 裴程程[1] 胡殊[1] 林书铭[1] 

机构地区：[1]沈阳出入境检验检疫局,辽宁沈阳110016

出　　处：《中国动物检疫》2012年第5期38-41,共4页China Animal Health Inspection

摘　　要：根据犬瘟热病毒(canine distemper virus,CDV)和犬腺病毒2型(Canineadenovirus type2,CAV-2)GeneBank登陆的基因序列各设计一对引物,经试验条件优化,建立了检测CAV-2的PCR方法和CDV的RT-PCR方法,CAV-2可扩增1108bp片段、CDV可扩增560bp的目的片段,特异性试验表明建立的方法只能特异扩增CAV-2和CDV,不能扩增犬细小病毒、犬冠状病毒、犬副流感病毒、犬钩端螺旋体和犬黄胆出血群、狂犬病病毒。敏感性试验表明,所建立的双重PCR方法可以扩增4.63ng总核酸量。在上述试验基础上继续优化条件建立了能同时检测CAV-2和CDV两种病毒的双重PCR检测方法,并用该方法检测来自辽宁、吉林等15家动物医院134份鼻液和眼眵样品,与商品化试纸条检测结果相比,本方法更加敏感和方便。研究结果表明,本实验建立的方法敏感、特异,适用于动物犬瘟热病毒和犬腺病毒2型的快速检测和鉴别诊断。A pair of PCR primers were designed according to sequence of CAV-2 and CDV respectively, which could amplify 1 108 bp fragment for CDV and 560 bp fragment for CAV-2. The products of PCR were proved to be specific.The sensitivity of PCR for CAV-2 and CDV showed that the method could amplify 4.63ng nucleic acid.The result of rudimentary application showed that the method was specific, sensitive, efficient, rapid and provided a new detecting method for the mixed infection with CAV-2 and CDV.

关 键 词：犬瘟热病毒 犬腺病毒2型 双重PCR 鉴别诊断 

分 类 号：S851.33[农业科学—预防兽医学]