检测猪伪狂犬病病毒gE抗体红细胞凝集试验的建立及应用

以纯化的抗人红细胞单链抗体(ScFv)—猪伪狂犬病病毒(PRV)gE蛋白双功能融合蛋白为抗原,建立了检测猪伪狂犬病毒gE抗体的红细胞凝集试验。利用方阵滴定试验筛选出最佳抗原工作浓度为55μg/mL,血清最佳稀释度为1∶20,作用时间15 min,与猪瘟(CSF)、猪细小病毒(PPV)、猪繁殖与呼吸综合征(PRRS)、猪乙型脑炎(JE)、猪布氏杆菌病(Brucellosis)阳性血清和PRV gE缺失疫苗接种的猪免疫血清均不出现红细胞凝集现象,与PRV标准阳性血清反应出现肉眼可见的凝集圈。与美国进口的PRV抗体检测gE-ELISA诊断试剂盒检测结果比较,50份猪血清的阴、阳性检出符合率均为100%。红细胞凝集试验检测方法具有操作简便、敏感性和特异性较高的特点,可用于PRV野毒感染的快速筛查。

The Development and Application of Erythrocyte Agglutination Assay for Detection of the Antibody to gE of Pseudorabies Virus in Porcine

Abstract：The erythrocyte agglutination assay for detection of the antibody to gE protein of pseudorabies virus in porcine was established by the anti- human red blood cell single chain fragment variable- pseudorabies virus gE bifunctional fusion protein as antigen. The optimal work concentration of antigen was 55 μg/mL. The dilution of sera was 1∶20. The reaction time of sera was 15 min. It revealed a negation reaction with positive sera of classical swine fever virus (CSFV), porcine parvovirus virus (PPV), porcine reproduction and respiratory syndrome virus (PRRS), Japanese encephalitis virus (JEV), Brucella and serum of swine inoculated by PRV gE deleted vaccines. It revealed a positive reaction with PRV standard positive serum. The total coincidence rate of the negative and positive reaction reached 100%. The results showed that the established erythrocyte agglutination assay has the advantages such as high sensibility and strong specificity and can be used to detect the porcine serum antibodies to gE protein of PRV.