猪伪狂犬病血清抗体gE-ELISA检测方法的建立

Establishment of the Indirect ELISA for the Detection of Viral Glycoprotein E Antibodies against Swine Pseudorabies

以纯化的猪伪狂犬病病毒gE蛋白为抗原建立了检测猪伪狂犬病血清抗体的间接gE-ELISA方法。最佳反应条件为,抗原包被浓度为1.7μg/mL,待检测血清1∶40稀释。与伪狂犬病阳性血清反应为阳性,与猪瘟、猪细小病毒病、猪繁殖与呼吸综合征(猪蓝耳病)、猪乙型脑炎、猪布氏杆菌病5种疾病阳性血清和猪伪狂犬病gE缺失疫苗接种的猪免疫血清及SPF猪阴性血清均无交叉反应。批间、批内试验变异系数分别不超过5%和9%。用该方法与Ingezim ELISA试剂盒和HerdChek ELISA试剂盒同时对172份血清进行了平行检测,总符合率分别达93.6%和83.7%。试验结果表明:猪伪狂犬病血清抗体间接gE-ELISA检测方法具有较高的敏感性和特异性,且重复性好,可用于猪伪狂犬病野毒感染猪的血清抗体检测。

The gE-ELISA was established by the recombinant protein as antigen, and all reaction conditions of this gE-ELISA were explored, the optimal coating concentration of antigen was 1.7 /μg/mL, the serum sample was diluted to 1：40. When sera collected from PRV infected pigs were detected by this gE-ELISA, the results were positive, and when pig sera which contained antibodies against hog holera virus, Japanese encephalitis virus, porcine reproductive and respiratory syndrome virus, porcine parvovirus and brucella were detected by this gE-ELISA,all results were negative. The CV between and within batches of detection were less than 5% and 9%. 172 pig sera samples were detected for the gE antibody by this indirect gE-ELISA, Ingezim ELISA kit and HerdChek ELISA kit, the conformation rates with the two kits was 93.6% and 83.7% respectively. All results showed that this assay had a good sensitivity, specificity and repeatability, it could be used for diagnosis of PRV infection.