MEV、ADV和CAV三种病毒多重PCR检测方法的建立  

Establishment of a Multiplex PCR for Detection of MEV,ADV and CAV

作　　者：罗彬[1] 易立[1] 王建科[1] 程世鹏[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：《动物医学进展》2009年第2期8-11,共4页Progress In Veterinary Medicine

基　　金：吉林省科技发展计划项目(20080213)

摘　　要：建立一种同时检测貂肠炎病毒(MEV)、貂阿留申病毒(ADV)和犬腺病毒(CAV)的多重PCR诊断方法。引用已有的CAV引物,并根据GenBank发表的MEV、ADV序列保守区域设计特异性引物进行PCR扩增,可同时得到扩增长度为795(MEV)、451(ADV)、1 019 bp(CAV)3条特异性片段,对猪细小病毒(PPV),犬瘟热病毒(CDV)进行PCR检测结果为阴性。各种模板、引物之间相互不构成干扰。敏感性试验证明,可以检测到模板中MEV101.5TCID50和CAV100.5TCID50的病毒含量,对ADV检测的敏感性更高。A novel multiplex PCR（mPCR）assay was developed and subsequently evaluated for its effectiveness in simultaneously detecting multiple viral infections of Mink enteritis virus （MEV）,Aleutian disease virus（ADV） ,Canine adenovirus（CAV）. Based on the reports and sequences published in GenBank, three pairs of specific primers were designed according to the conservative compass. Three pairs of specific primers for each of these virus genomes MEV/ADV/CAV were amplified to detect 795 bp（MEV）,451 bp （ADV）, 1 019 bp（CAV）,respectively. No band was amplified from PPV and CDV in the specificity test. In the sensitivity test, it can detect the virus MEV 10^2.5 TCID50, ADV 1 TCID50, CAV far more than 10^1.5 TCID50 in template. The multiplex PCR has potential to apply in diagnosis of combined infections in these three diseases.

关 键 词：多重PCR 貂肠炎病毒 貂阿留申病毒 犬腺病毒 

分 类 号：S852.65[农业科学—基础兽医学]