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

摘要

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 inoculaled with tine virus. The N gene of isolate shared 95. 5% to 96. 9% nucleo-tide homology with that of vaccine stainsfOnderstepoort and Snyder Hill) .and 97. 2% to 99. 0% nucleotide bomology 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.%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、Z J7、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.