间接ELISA检测抗禽白血病病毒抗体方法的建立  

Development of an Indirect ELISA for the Detection of Antibodies Against Avian Leukosis Virus

作　　者：仇钰[1] 秦爱建[1] 钱琨[1] 金文杰[1] 胡序明[1] 沈海玉[1] 

机构地区：[1]扬州大学江苏省动物预防医学重点实验室,江苏扬州225009

出　　处：《中国家禽》2010年第22期16-20,共5页China Poultry

基　　金：国家行业专项(200803019);江苏省高校自然科学重大基础研究项目(07KJA23021);NSFC-广东联合基金项目(U0831002)

摘　　要：禽白血病病毒(Avian leukosis virus,ALV)是禽白血病的病原,可引起鸡的免疫抑制和肿瘤。净化种鸡群是控制ALV的主要方法之一。本研究将ALV的p27基因克隆到表达载体pGEX-6P-1,在大肠杆菌中获得了高效表达,以纯化后的p27-GST融合蛋白为抗原包被,经过条件优化,建立了检测鸡血清中抗ALV抗体的间接ELISA方法。与IFA检测结果比较,该方法比IDEXX ELISA试剂盒有更高的符合率。可用于禽白血病病毒感染根除的大规模检测,并具有低成本、易操作的特点,能同时检测到针对ALV所有亚群的抗体。Avian leukosis virus （ALV）is associated with tumor development and immunosuppression in pouhry industry. Eradication of virus infection at the primary breeder level is the principal method for controlling ALV infection in chickens. An indirect ELISA （iELISA） method with the prokaryotic expressed and affinity-purified viral capsid protein antigen p27 was developed for the detection of AI,V-speeific antibodies in chicken sera. The protocol of iELISA was optimized and the method showed more sensitive and specific than the commercial IDEXX ALV antibody test kit when immunofloureseent assay （IFA） was used as a reference. The main advantage of this method is the purified fusion protein to detect antibodies against all ALV subgroups. Data in this paper indicated that the developed iELISA kit was cheap,easy to perform and could be used as a confirmatory test for the presence of anti-ALV antibodies on a large scale.

关 键 词：禽白血病病毒 P27 原核表达 间接ELISA 

分 类 号：S852.65[农业科学—基础兽医学]