基于非洲猪瘟病毒特异性单克隆抗体建立的阻断ELISA方法  

Establishment of a Blocking ELISA Method Based on AfricanSwine Fever Virus Specific Monoclonal Antibody

作　　者：董春娜 张蕾 李静 李鹏宇 齐鹏 肖进 DONG Chun-na;ZHANG Lei;LI Jing;LI Peng-yu;QI Peng;XIAO Jin(China Animal Husbandry Industry Co.,Ltd.,Ministry of Agriculture Key Laboratory of Veterinary Biologics and Drugs,Veterinary Peptide Vaccine Design and Preparation of Engineering and Technology Center of Beijing,Beijing 100095,China)

机构地区：[1]中牧实业股份有限公司农业农村部兽用生物制品与化药重点实验室北京市兽用多肽疫苗设计与制备工程技术中心,北京海淀100095

出　　处：《中国兽医杂志》2023年第3期36-40,共5页Chinese Journal of Veterinary Medicine

摘　　要：为了建立一种能够检测非洲猪瘟病毒(ASFV)特异性抗体的单抗阻断ELISA方法,本试验首先构建了非洲猪瘟病毒p30蛋白的原核表达载体,进而制备并筛选出1株能特异性识别p30蛋白的高亲和力单克隆抗体,采用昆虫杆状病毒表达系统Bac-to-Bac进行该单克隆抗体的表达。以p30重组蛋白作为包被抗原,辣根过氧化物酶(HRP)标记的单克隆抗体作为阻断抗体建立了一种非洲猪瘟病毒特异性抗体的检测方法,并验证其敏感性、特异性以及与商品化试剂盒的符合率。结果显示,该方法敏感性为100%,特异性为100%,与商品化试剂盒比较,符合率为98.7%。结果表明,本试验基于非洲猪瘟病毒特异性单克隆抗体建立的阻断ELISA方法敏感性好、特异性强,结果准确可靠,可用于非洲猪瘟病毒抗体检测。This study aimed to develop a monoclonal antibody blocking ELISA method for detecting specific antibodies to African swine fever virus(ASFV).A prokaryotic expression vector for ASFV p30 protein was constructed,and then a monoclonal antibody with high affinity was prepared and screened for specific recognition of p30 protein.The monoclonal antibody against p30 protein was expressed by Bac-to-Bac insect baculovirus expression system.Using the recombinant p30 protein as the coating antigen and horseradish peroxidase(HRP)labeled monoclonal antibody as the blocking antibody,an ELISA for detection of ASFV-specific antibody was established and its sensitivity,specificity and coincidence rate with commercial kit were assessed.The results showed that the ELISA had sensitivity and specificity of 100%,and coincidence rate of 98.7%compared with the commercial kit.Thus,the blocking ELISA based on specific monoclonal antibody of ASFV provides a sensitive,specific and accurate method for the detection of ASFV antibodies.

关 键 词：非洲猪瘟病毒 p30蛋白 阻断ELISA 

分 类 号：S855[农业科学—临床兽医学]