›› 2009, Vol. 36 ›› Issue (3): 153-156.

• 疾病防治 • 上一篇    下一篇

貉源犬瘟热病毒的分离鉴定

刘双翼,王进轶,王延涛,刘合义,侯喜林   

  1. (黑龙江八一农垦大学动物科技学院,大庆 163319)
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2009-03-20 发布日期:2009-03-20
  • 通讯作者: 侯喜林

Isolation and Identification of Canine Distemper Virus from Raccoon Dog

LIU Shuang-yi ,WANG Jin-yi,WANG Yan-tao,LIU He-yi,HOU Xi-lin   

  1. (Animal Science College,HLJ August First Land Reclamation University,Daqing 163319, China)
  • Received:1900-01-01 Revised:1900-01-01 Online:2009-03-20 Published:2009-03-20
  • Contact: HOU Xi-lin

摘要: 采集大庆某貉养殖场病貉的病料,处理后接种于非洲绿猴肾细胞(Vero)进行病毒分离。对分离毒株进行了中和试验、血凝试验、理化性质的鉴定,并用反转录-聚合酶链式反应(RT-PCR)检测感染细胞中的病毒核酸。病料接种 Vero 细胞 72 h 后产生明显的细胞病变(CPE);病毒分离株可以被犬瘟热病毒阳性血清中和,中和效价为 1∶25;分离株对氯仿、乙醚敏感,对酸和热抵抗力弱,可以凝集鸡红细胞,其凝集作用可被犬瘟热病毒阳性血清所抑制; RT-PCR检测病毒细胞培养液,扩增出的片段长287 bp,与预期设计的长度相同,经测序发现与MS01株的同源性为99%。结果表明,分离的病毒株为犬瘟热病毒,命名为CDV-DQ株。

关键词: 貉; 犬瘟热病毒; 分离; 鉴定

Abstract: Raccoon dog specimen from daqing were collected for virus isolation in vero cell culture. The isolated virus was confirmed by a series of systemic identification, such as serology test ,physics and chemistry characteristics and molecular virology test.The results showed that the specimens could cause cytopathogenic efect (CPE) in vero cell after subculturing 72 h. The isolated virus could be neutralized with positive canine sera of CDV with the neutralization titer of 1∶25.The agglutination titer of isolated virus was 1∶16 for the chicken erythrocytes. The isolated virus was sensitive for chloroform, aether and acid. It was completely inactivated at 60 ℃ within 1 h. A DNA fragment of 287 bp was amplified from infected vero cells by RT-PCR.The agreement of N gene fragment sequence between the isolated virus and CDV MS01 from GenBank was 99%.From the results described above,the isolated virus was CDV and designated as CDV-DQ.

Key words: raccon dog; canine distemper virus; isolation; identification

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