犬瘟热病毒的分离与鉴定

Isolation and identification of a strain of canine distemper virus

本试验收集了疑似犬瘟热病毒(CDV)感染病犬的粪便拭子,预处理后接种于非洲绿猴肾细胞(Vero)进行病毒分离培养。对分离出的病毒进行毒力测定、电镜形态学观察、特异性基因鉴定和N基因全长扩增、间接免疫荧光(IFA)检测。结果显示,在Vero细胞盲传至第4代,90%以上细胞出现稳定的细胞毒性改变(CPE);病毒粒子有囊膜,形状不规则;RT-PCR成功扩增出特异性基因和N基因。IFA结果显示,在细胞胞质中可观察到特异性绿色荧光。结果表明,本试验成功分离并鉴定出1株宁夏地区CDV毒株。

A swab of dog＇s feces,which were suspected infected by canine distemper virus（CDV）,was collected.The suspicious materials were pretreated and inoculated in Vero cell line to isolate virus.Then toxicity measurement,electron microscopy morphology test,specific identification of PCR,N gene amplification of PCR and indirect immunofluorescent assay（IFA） were carried out on the isolated virus.The results showed that more than 90% cells have stable toxicity changes after blindly passage to the fourth generation.Virus particles have envelopes and irregular shapes,the specific gene and N gene were amplified successfully.Specific green fluorescence in the cell cytoplasm can be observed by IFA.A CDV epidemic strain was isolated and identified successfully in Ningxia.