猪非典型瘟病毒的基因组特征分析及荧光定量RT-PCR检测方法的建立  

Genomic characterization and development of real-time RT-PCR for the detection of atypical porcine pestivirus

作　　者：许古明[1] 葛士坤 张凯照 刘健新[1] 任旭皎 崔灝 罗畅[1] 宁章勇[1] 

机构地区：[1]华南农业大学兽医学院,广东广州510642

出　　处：《中国兽医科学》2017年第11期1357-1362,共6页Chinese Veterinary Science

摘　　要：最近的研究表明,猪非典型瘟病毒(APPV)在我国猪群流行,并造成了严重的危害。本研究应用RT-PCR方法对APPV GD3株进行了全基因组扩增、测序,并进行了全基因组及其部分编码基因的遗传演化分析;根据APPV基因组编码的结构蛋白E2的基因序列,构建了APPV的荧光定量RT-PCR检测方法。APPV GD3株的全基因组序列与德国、美国、荷兰和西班牙毒株在不同的进化分支上,同源性为83.1%~83.4%;与中国毒株GD1、GD2和GD的同源性分别为99.5%、99.6%和83.4%。GD3株编码的E2、Npro和Erns基因与国内外现有毒株的同源性分别为83.1%~99.7%、80.2%~99.8%和81.9%~99.5%。本研究建立的荧光定量RTPCR检测方法特异性强、最低检测浓度为10 copes/L、操作简单、耗时短,具有良好的兽医临床应用前景。The most recent researches showed that atypical porcine pestivirus（APPV） was prevalent in swine herds in China and caused serious losses. In this research,the whole genome of APPV GD3 strain was amplified by RT-PCR,and the genome sequence and its partial coding genes were genetically analyzed.According to the structural protein E2 gene sequence encoded by APPV genome,a real-time RT-PCR method was established for detection of APPV. The whole genome sequence of APPV shared from 83.1%to 83.4%homology with German,American,Dutch and Spanish strains and APPV located at a different branch.The homologies between GD3 and GD1,GD2 and GD strains were 99.5%,99.6% and 83.4%,respectively.The homology of E2,Nproand Ernsencoded by GD3 strains of APPV with currently available strains were from83.1% to 99.7%,80.2% to 99.8% and 81.9% to 99.5%,respectively.The real-time RT-PCR method established in this study was highly specific,simple,time-saving and the detection limit was 10 copies/L.It will be well used in veterinary clinical detection.

关 键 词：猪非典型瘟病毒 序列分析 荧光定量RT-PCR 

分 类 号：S852.723[农业科学—基础兽医学]