鸡传染性法氏囊病毒蚀斑克隆纯化CV01株生物学特性研究  

Characterization of a strain of infectious bursal disease virus(CV01) by plaque purification

作　　者：揭鸿英 朱亚露 毕志香 赵阳 何家惠 于漾 侯继波 

机构地区：[1]江苏省农业科院国家兽用生物制品工程技术研究中心,江苏南京210014 [2]江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009

出　　处：《畜牧与兽医》2018年第2期86-90,共5页Animal Husbandry & Veterinary Medicine

基　　金：公益性行业(农业)科研专项(201303046)

摘　　要：通过蚀斑克隆技术,对野外分离的鸡传染性法氏囊病毒（IBDV）进行纯化后,筛选到1株病毒含量高、传代稳定的蚀斑纯化毒CV01株。生物学特性分析发现,IBDV能在鸡胚成纤维细胞（DF-1）上形成大小不同的蚀斑,且蚀斑大小与病毒的毒力有一定的相关性,大蚀斑的病毒含量高（10^8.0TCID50/m L以上）,小蚀斑的病毒含量低（10^7.0l-10^8.0TCID50/m L）。蚀斑克隆VP2基因序列分析显示大蚀斑克隆毒具有IBDV超强毒株的特性。选病毒含量高（10^8.5TCID50/m L）的大蚀斑CV01进行进一步试验,结果发现CV01对10日龄SPF鸡胚的毒力达到10^6.7/0.2 m L以上,能致40%的30日龄SPF鸡感染。蚀斑克隆毒的免疫原性好,血清中和抗体水平高达15.6 log2,能100%抵抗超强毒株的攻击,表明该纯化株适合用于疫苗生产。A strain of IBDV（CV01）,with high virus titer and stability,was isolated from a cell. The strain of IBDV was adapted by plaque purification assay and its biological characteristics were studied. In the continuous cell line,DF-1,a plaque of different sizes was found after IBDV inoculation. It was also found that there was a correlation between the size of the plaque and the virulence of the virus. The virus titer of the large plaque was over 10^8. 0 TCID50/ml,while the virus titer of the small plaque was between 10^7. 0 TCID50/ml and 10^8. 0 TCID50/ml. Sequence analysis based on the nucleic acid identities of VP2 revealed that the virus on the large plaque shared the highest similarity with vv IBDV,indicating that the virus strain on the large plaque belonged to vv IBDV. A strain of IBDV from the large plaque,with the highest virus titer（10^8. 5 TCID50/ml）,was named CV01 and was selected for further study. The result of the inoculation test showed that the titer of the virus on the 10-day SPF embryo could reach as high as 10^6. 7/0. 2 ml and the infection rate on the 30-day chicken was 40%.CV01 also showed good immunogenicity and the titer for the neutralizing antibody of the serum was 15. 6 log2,which could provide full protection for the chicken from vv IBDV infection,suggesting that CV01 had a potential of application to research on and development of vaccine against IBDV in the future.

关 键 词：鸡传染性法氏囊病毒 蚀斑 纯化 生物学特性 

分 类 号：S852.6[农业科学—基础兽医学]