貉源犬瘟热病毒的分离鉴定及其H基因的序列分析

Isolation and identification of canine distemper virus from raccoon dog and sequence analysis of its gene H

为了研究我国貉源犬瘟热病毒(CDV)的流行、遗传变异性,试验从某养殖场选取5只疑似患犬瘟热病死貉的肝脏、肺脏、脾脏、肠等脏器,利用Vero细胞分离出1株病毒,通过免疫荧光、RT-PCR等方法鉴定为CDV,命名为HLJ(11),利用其RNA为模板,采用RT-PCR扩增血凝素(H)基因,进行序列测定与分析。结果表明:分离株HLJ(11)的H基因序列与HeB(07)1、SD(09)3、SD(07)1同源性最高,分别为99.9%、99.7%、99.6%,与Strain CDV 3、Convac vaccine strain的同源性最低,为91.4%;遗传进化分析显示,分离株HLJ(11)为Asia-Ⅰ型。

To study the prevalence and variability of canine distemper virus （CDV） from raccoon dog in China, the livers, lungs, spleens, intestines and other organs were collected from five dead raccoon dogs suspected with CDV infection in a certain farm, and then a strain of virus was isolated using vero cells culture. The virus was identified to be a canine distemper virus by immunofluorescence, RT - PCR and other meth- ods, which was named HLJ （ 11 ） . The H gene of the isolated strain was amplified by RT - PCR, and then was cloned and sequenced, taking the RNA from the isolate HLJ （ 11 ） as a template. The results showed that the homologies of nucleic acid sequences for the H gene from HLJ （ 11 ） with HeB （ 07 ） 1, SD （09） 3 and SD （ 07 ） 1 were high （ 99.9 %, 99.7 %, 99.6 %, respectively）. The homologies of nucleic acid sequences with Strain CDV 3 and Convac vaccine strain were low（91.4% ）. The phylogenetic tree showed that the isolated strain belonged to Asia - I.