犬瘟热病毒的分离

Isolation of canine distemper virus

对流行病学调查、临床症状检查和血清学(ELISA)检测为阳性的自然发病犬,取肠内容物为病料,采用同步培养方法分别接种于非洲绿猴肾细胞系(Vero)、犬肾细胞系(MDCK)进行病毒的分离。用多聚酶链式反应(RT-PCR)检测感染细胞中的病毒核酸。结果表明,病料接种Vero细胞、MDCK细胞均产生明显的细胞病变(CPE);用RT-PCR技术检测病毒感染的细胞培养液,扩增出特异性的产物带,扩增出的片段大小分子长为760bp,与预期设计的长度相同。用Vero细胞同步培养从肠内容物中分离出的病毒命名为犬瘟热病毒GZ1株;用MDCK细胞同步培养从另一只犬的肠内容物中分离出的病毒命名为犬瘟热病毒GZ2株。

Canine distemper virus（CDV） was isolated from dead dogs which were suspected as canine distemper through epidemiologic investigation ,clinical symptom detection and ELISA diagnosis. The intestinal contents were collected from affected dogs for virus isolation in Vero monkey kidney（Vero） cells and Madin- Darby canine kidney（MDCK） cell culture. The virus RNA was detected by reverse transcription-polymerase chain reaction （RT- PCR）. The results showed that the affected material could multiply rapidly in Veto cells, MDCK cells and caused cytopathogenic effect（CPE）. A DNA fragment of 760bp was amplified from infected Vero, MDCK cells by RT - PCR. A virus strain was isolated from infected Vero cells and designated as CDV - GZ1. The other was isolated from infected MDCK cells and designated as CDV - GZ2.