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

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.