犬细小病毒和犬冠状病毒双重PCR方法的建立及应用  

Establishment and Application of Duplex PCR for CPV and CCV Detection

作　　者：林颖[1] 耿庆华[1] 付海滨[1] 李俊环[1] 闫明媚[2] 李成镛[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 [2]辽宁省动物疫病预防控制中心,辽宁沈阳110161

出　　处：《动物医学进展》2011年第10期71-74,共4页Progress In Veterinary Medicine

摘　　要：根据GenBank中犬细小病毒(CPV)和犬冠状病毒(CCV)基因序列各设计了一对引物,经试验条件的优化,建立了检测CPV的PCR方法和CCV的RT-PCR方法,扩增目的片段大小分别为337bp和852bp。在此基础上建立了检测CPV和CCV双重PCR方法。该双重PCR方法能特异的扩增CPV和CCV,且分别扩增出1.08ng和3.2ng总核酸量。通过检测96份临床样品,对双重PCR方法和胶体金试纸条方法进行了对比试验。结果表明,建立的双重PCR方法快速、敏感、特异性高,可以用于CPV和CCV鉴别检测。One pair of PCR primer were designed according to sequence of Canine parvovirus（CPV） and Canine coronavirus（CCV） from GenBank respectively,which could amplify 337 bp fragment for CPV and 852 bp fragment for CCV.The products of PCR were proved to be specific.The sensitivity of PCR of CPV and CCV showed that the method could amplify 1.08 ng and 3.2 ng nucleic acid.The result of rudimentary application showed that the method was specific,sensitive,efficient,rapid,and was a new detecting method for the mixed infection of CPV and CCV,which also can differentiate CPV and CCV.

关 键 词：犬细小病毒 犬冠状病毒 双重PCR 

分 类 号：S852.659[农业科学—基础兽医学] S852.659[农业科学—兽医学]