犬瘟热病毒山东株的分离及N蛋白基因的克隆与序列分析

本试验采用Vero细胞从临床疑似患犬瘟热的犬组织病料中分离出一株病毒,通过观察细胞病变,提取细胞培养物RNA并扩增克隆分析N蛋白基因部分序列进行了鉴定。结果表明,该毒株接种于Vero细胞后,出现了典型的细胞病变;对接种病料后的Vero细胞培养物提取总RNA进行RT-PCR扩增,得到了287 bp的目的片段;序列分析表明该分离株与犬瘟热病毒弱毒(Onderstepoort和Snyder Hill)的同源性为95.5%~96.9%,与标准强毒株(A75/17和2544/Han95)及野毒株(XJ-4、ZJ7、98-2646等)的同源性较高,为97.2%~99.0%,证明该毒株为犬瘟热病毒野毒株,命名为QD10。

Isolation,Cloning and Analysis of Nucleoprotein Gene of Canine Distemper Virus in Shandong

A strain of canine distemper virus(CDV) was isolated from the domestic dog in Shandong province,the virus was identified by cell pathological effect observation and RT-PCR.The results showed that typical pathological changes appeared on Vero cells inoculated with the treated pathological material.A 287 bp fragment of nucleoprotein(N) gene was amplified from RNA extraction of Vero cell culture inoculated with the virus.The N gene of isolate shared 95.5% to 96.9% nucleotide homology with that of vaccine stains(Onderstepoort and Snyder Hill),and 97.2% to 99.0% nucleotide homology identity with that of virulent strains(A75/17 and 2544/Han95) and field strains(XJ-4,ZJ7,98-2646 and so on),the phylogenetic tree also suggested that the isolate had close relationship with field strains,it indicated that the isolate was a CDV field strain,which was named QD10.