一株禽腺病毒4型病毒分离与鉴定  

Isolation and Identification of a Strain of FAV4

作　　者：李坤林[1] 于新友 LI Kunin;YU Xinyou(Shandong Binzhou Animal Science&Veterinary Medicine Academy,Binzhou 256600,Shandong China;Shandong Lvdu Bio-Industry Co.,Ltd.,Binzhou 256600,Shandong China)

机构地区：[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

出　　处：《饲料博览》2023年第5期56-60,共5页Feed Review

基　　金：山东省现代农业产业技术体系家禽创新团队计划资助项目(SDAIT-11-16)。

摘　　要：为了对某肉鸡场暴发的疾病进行确诊,对送检的病死鸡进行解剖、细菌分离和荧光PCR检测,并进一步进行鸡胚半数致死量(ELD50)、动物回归试验和组织灭活疫苗研制。结果显示:病死鸡剖检可见肝脏肿大、出血,心包胶冻样积液,肾脏肿大,病料未分离到细菌。PCR检测禽腺病毒4型(FAV4)阳性,用处理液进行病毒分离,成功分离到一株FAV4病毒,扩增所分离病毒hexon基因部分序列,测序结果与GenBank上的FAV4hexon基因(登录号:EF554403)相似性为100%。分离病毒盲传3代,测定的ELD50为104.5/0.2mL,以0.2mL·只^(-1)的剂量胸部肌肉注射30日龄SPF鸡5只,4 d内鸡只全部死亡,病变与临床剖检病死鸡相似。取病死鸡肝脏制备组织灭活疫苗,免疫SPF鸡,能抵抗所分离FAV4病毒攻击。研究成功分离鉴定出一株FAV4病毒,并进行初步研究,丰富了毒种库,为FAV4病毒疫苗研制奠定了基础。In order to diagnose the disease broken out in a broiler farm,the sick and dead chickens were submitted for dissecting,isolating bacteria,detecting real time PCR,and further to analyze the embryo median lethal dose(ELD50)of chickens,animal regression test and tissue inactivated vaccine development.The autopsy results of the dead chicken showed liver enlargement and bleeding,pericardial jelly like fluid accumulation,kidney enlargement,and no bacterium was isolated from the diseased material.The PCR detection of fowl adenovirus type 4(FAV4)was positive,and a strain of FAV4 virus was successfully isolated using the treatment solution.The partial sequence of the isolated virus hexon gene was amplified,and the sequencing results showed 100%similarity with the FAV4 hexon gene(login number EF554403)in GenBank.After blind passaging for three generation,the ELD50 of virus was 104.5/0.2 mL.Virus was injected into the chest muscle of five 30-day-old SPF chickens at a dose of 0.2 mL each one,all the chickens died within 4 days,and the pathological changes were similar to those of clinical dead chickens.Prepare tissue inactivated vaccines from the liver of dead chickens,and immunize SPF chickens to resist the attack of the isolated FAV4 virus.The study successfully isolated and identified a strain of FAV4 virus,conducted preliminary research,enriched the virus seed bank and laid the foundation of the development of FAV4 virus vaccines.

关 键 词：禽腺病毒4型 分离 鉴定 

分 类 号：S858[农业科学—临床兽医学] S851.33[农业科学—兽医学]