我国鸡传染性支气管炎病毒LD3/01/07的分离鉴定  

Isolation and Identification of an Avian Infectious Bronchitis Virus Strain LD3/01/07 from China

作　　者：刘胜旺[1] 孔宪刚[1] 刘玉芬[2] 杜恩岐[3] 谷守林[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨150001 [2]东北农业大学预防兽医学 [3]西北农林科技大学

出　　处：《中国预防兽医学报》2003年第2期121-127,共7页Chinese Journal of Preventive Veterinary Medicine

基　　金：黑龙江省政府博士后科研启动金资助项目(LRB_KY0 10 4 5 )

摘　　要：将我国黑龙江省某鸡场发病症状及病理变化疑似鸡传染性支气管炎的濒死病鸡肾脏组织加PBS缓冲液后研磨 ,- 70℃ / 37℃反复冻融三次 ,离心并进行无菌过滤后接种 10日龄SPF鸡胚。 72小时取尿囊液进行电镜观察并用 1%胰酶处理 ,观察对鸡红细胞的凝集情况。结果发现尿囊液中存在典型的冠状病毒粒子 ,而且用胰酶处理过的尿囊液能够凝集SPF鸡的红细胞 ,初步鉴定为鸡的传染性支气管炎病毒 (InfectiousBronchitisVirus ,IBV)。将该病毒尿囊液在 10日龄SPF鸡胚上连续传 8代 ,通过病毒对鸡胚的致病作用、病毒在电镜下的形态特征、病毒在鸡胚中的增殖动态以及病毒凝集鸡红细胞的特性等对该毒株进行研究。结果表明 :该毒株的第一代尿囊液中病毒的滴度很高 ,可达到 8log2 ,且胚体表现轻度出血 ,但实质器官未见肉眼可见的病变 ;第二代病毒即可致胚体严重病变 ;病毒在鸡胚中随着接种病毒后时间的延长 ,其滴度逐渐增高 ;经 1%胰酶处理后的各代病毒尿囊液可凝集鸡红细胞。鸡胚的第三代病毒尿囊液 (命名为LD3/ 0 1/0 7)分别接种 1日龄和 30日龄的SPF鸡 ,发现对不同日龄的鸡表现不同的致病性 ,对 1日龄接种鸡和同笼饲养的对照鸡致病力高 ,发病率和致死率均为 10 0 % ,(接种和同居感染各 4只 ,均死亡 ) ,30日龄接种和同居感?A field strain of infectious bronchitis virus (IBV) was isolated from chicken kidney of the affected young layer. Biological characteristics of the virus were studied by virus pathogenicity to chicken embryo,morphological observation by electron_microscope,hemagglutination activity and virus pathogenicity to SPF chickens. It was demonstrated that allantoic fluid of the first passage virus produced mild pathogenisis to chicken embryos and the gross lesions caused by second passage virus were obvious. The virus showed typical shape shared by members of Coronaviridae under electron_microscope and could hemagglutinate chicken red blood cells after treatment with 1% trypsin. The morbidity and mortality of the third passage virus (LD3/01/07) to 1_day_old chickens were 100% and to 30_day_old chickens were 100% and 20% respectively. Moreover, the gross lesions in kidney caused by LD3/01/07 were distinct and this showed that the virus was nephropathogenic avian infectious bronchitis virus. In addition, N gene of LD3/01/07 shared the typical molecular characteristics of all IBV N genes.

关 键 词：鸡传染性支气管病毒 LD3/01/07 分离鉴定 

分 类 号：S858.312.6[农业科学—临床兽医学]