中国畜牧兽医

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水貂犬瘟热病毒LD-1株的分离与鉴定

庄金秋13,梅建国1,王金良1,张倩2,沈志强12,丁壮3   

  1. (1.山东省滨州畜牧兽医研究院,山东滨州 256600; 2.山东绿都生物科技有限公司,山东滨州 256600;3.吉林大学畜牧兽医学院,吉林长春 130062)
  • 收稿日期:2013-10-24 出版日期:2014-04-20 发布日期:2014-05-27
  • 通讯作者: 沈志强。E-mail:bzshenzq@vip.sina.com 丁壮。E-mail:dingzhuang@jlu.edu.cn
  • 作者简介:庄金秋(1978—),女,山东人,硕士生,助理研究员,研究方向:细胞培养和动物用病毒疫苗研制。
  • 基金资助:

    山东省博士后创新项目(201303027)。

Isolation and Identification of Mink Canine Distemper Virus LD-1 Strain

ZHUANG Jin-qiu1,3, MEI Jian-guo1, WANG Jin-liang1, ZHANG Qian2, SHEN Zhi-qiang1,2, DING Zhuang3   

  1. 1.Shandong Binzhou Animal Science & Veterinary Medicine Academy, Binzhou 256600, China;2.Shandong Lvdu Bio Sciences & Technology Co.,Ltd., Binzhou 256600, China;3.College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China)
  • Received:2013-10-24 Online:2014-04-20 Published:2014-05-27

摘要: 取山东某貂场疑似犬瘟热病毒(canine distemper virus,CDV)感染的水貂肝脏等组织病料,通过RT-PCR检测呈CDV阳性,且无水貂细小病毒存在,将病料接种原代CEF细胞、传代系Vero细胞和DF1细胞3种细胞进行病毒分离,通过优化细胞培养条件,最终在Vero细胞上传代培养成功,出现露珠状典型细胞病变(CPE)。分离毒应用RT-PCR、PCR产物测序、抗体中和试验(SN)、间接免疫荧光试验(IFA)及病毒包涵体检查等多种方法进行了CDV的鉴定,结果显示CDV阳性,表明分离到的病毒为水貂CDV,并将其命名为CDV LD-1株。

关键词: 犬瘟热病毒; 病毒分离; 鉴定

Abstract: Canine distemper virus (CDV) was detected positive by RT-PCR from a suspected CDV infected mink liver tissue. No mink parvovirus was detected from it. The virus was inoculated to 3 kinds of cells including primary CEF cells, Vero cells and DF1 cells for virus isolation. It was successfully isolated from Vero cells through the passage culture by optimizing the cells culture condition, and showed the dew shape typical CPE. The isolated virus was identified by RT-PCR, sequencing of the PCR products, serum neutralization test, indirect immunofluorescence test, virus inclusion body examination and other methods. The results showed that CDV was positive. It was proved that the isolated virus was a mink CDV, named CDV LD-1 strain.

Key words: canine distemper virus; virus isolation; identification