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

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

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287bp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。

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 ap- peared on Vero cells inoculated with the treated pathological material. A 287 bp fragment of nucleoprotein（N） gene was ampli- fied from RNA extraction of Vero cell culture inoculated with the virus. The N gene of isolate shared 95.5% to 96.9% nucleo- tide 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, ZJT, 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.