猪伪狂犬病病毒单抗双夹心ELISA的构建  

Estabilishment of monoclonal antibodies double sandwich ELISA detecting porcine pseudorabies virus

作　　者：祁小乐[1] 崔保安[1] 牛春玲[1] 王莉娟[1] 刘占通[1] 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450002

出　　处：《河南农业大学学报》2004年第4期387-393,共7页Journal of Henan Agricultural University

基　　金：国家"十五"重大攻关项目(2001BA804A30-11)

摘　　要：建立了检测猪伪狂犬病病毒(Pseudorabiesvirus,PRV)的单克隆抗体双夹心酶联免疫吸附试验(Enzyme linkedimmunosor bentassay,ELISA)方法,为该病的快速诊断奠定了基础.作者对其组装条件进行了筛选,认为兔抗PRVIgG最佳包被质量浓度为4.59mg·L-1;抗PRV的单克隆抗体(Monoclonalantibodies,McAb)的最佳质量浓度为6.45mg·L-1;最佳封闭剂为10g·L-1BSA(牛血清白蛋白),封闭时间为60min,制定了科学的结果判定标准,最低病毒检出量为200μg·L-1.与病毒分离、动物试验和中和试验相比较,该方法特异、灵敏、可靠、方便.The double sandwich enzyme-linked immunosorbent assay (ELISA) for detection of porcine Pseudorabies virus (PRV) was developed,which laid the basis of the diagnosis of the disease.The best coatinging quality concentration of the rabbit-anti-PRV IgG was 4.59 mg·L^(-1).The best quality concentration of McAb was 6.45 mg·L^(-1).(10 g·L^(-1) BSA-PBST) was selected as the optimal blocking reagent,and the optimal blocking time is 60 min.The scientific criterion of determination was made.The sensitivity of the double sandwich ELISA was 200 μg·L^(-1).The assay was specific,sensitive,reliable,and convenient.

关 键 词：猪 伪狂犬病病毒 双夹心ELISA 单克隆抗体 诊断 组装条件 

分 类 号：S858.28[农业科学—临床兽医学]