犬瘟热病毒NT株分离鉴定、全基因测序与分析

Isolation,Identifi cation and Whole Gene Sequencing of Canine Distemper Virus NT Strain

为分析我国犬瘟热病毒的流行性特点及阐明该毒株的基因特征与遗传进化情况。本研究用表达SLAM受体的犬源Vero细胞系对犬瘟热病毒分离培养,并从南通、安徽两地收集的疑似犬瘟热病料(肺,脾)中分离出一株犬瘟热病毒,运用RT-PCR、病毒的半数感染量(TCID50)等方法对分离株进行鉴定,从而确定分离株为犬瘟热毒株,命名为CDV-NT-1。利用RT-PCR方法分段扩增此分离株的全基因组cDNA,用DNAStar软件比较CDV-NT-1分离株与GenBank上其他典型CDV毒株的同源性,并用Mega7.0软件进行系统进化分析。结果显示:CDV-NT-1株全基因组序列长度为15544 bp;CDV-NT-1与野毒株的同源性在97.6%~98.5%,而与疫苗株Snyder Hill、Phoca、Onderstepoort的同源性只有92.1%,CDV-NT-1与Asia-I位于同一分支,与疫苗株处于不同的分支,属于Asia-1型。本研究结果为CDV的研究奠定了基础。

To analyze the epidemic characteristics of Canine distemper virus in China and genetic evolution,suspected distemper fever materials(lungs and spleens)were collected from NanTong andAnHui for inoculation onto Vero cells.As a result,a strain of Canine distemper virus was isolated and designated as CDV-NT-1.The full-genome of the isolate CDV-NT-1 was amplified by RT-PCR for comparison with other typical CDV strains in GenBank by DNAStar software and phylogenetic analysis by Mega7.0 software.The sequence length of CDV-NT-1 was 15544 bp,and its homology was 97.6%-98.5%with the wild strain but only 92.1%with the vaccine strains Snyder Hill,Phoca and Onderstepoort.The phylogenetic analysis showed that CDV-NT-1 was located in the same branch as Asia-I,which was different branch from the vaccine strain and laid the foundation for the future study of CDV.