乙型脑炎病毒（JEV）ELISA检测方法的建立

目的建立猪乙型脑炎病毒（JEV）抗体ELISA检测方法。方法培养BHK21细胞,接种JEV病毒,制备BHK21正常抗原和JEV特异抗原,滴定酶结合物和抗原最佳工作浓度,并进行精密性、敏感性、稳定性、特异性实验。结果正常、特异抗原和酶结合物最佳工作浓度分别为0.2μg/mL、10μg/mL和1∶20000;正常、特异抗原批内变异系数分别为8.3%和6.4%,批间平均变异系数分别为9.7%和11.5%;检测灵敏度为1∶1280;与猪瘟病毒（CSFV）、猪细小病毒（PPV）均无交叉反应。稳定性试验相对偏差小于25%。结论建立的ELISA方法重复性、稳定性好,特异性、敏感性强。可用于猪JEV抗体的检测。

Establishment of an ELISA Technique in Detecting Japanese Encephalitis Virus Antibody

Objective To establish an ELISA detection method for Japanese encephalitis virus（JEV） antibody.Methods BHK21 cells were cultured and JEV virus was vaccinated.The normal BHK21 antigen and JEV-specific antigen were prepared.The optimal working concentration of enzyme conjugate and the normal or specific antigens were titrated,and the accuracy,sensitivity,stability and specificity were tested.Result The best working concentrations of normal and specific antigens and the enzyme conjugate were 0.2 μg/mL,10 μg/mL and 1 ∶ 20000,respectively.The intra-assay coefficient of variation of normal antigen and special antigen was 8.3% and 6.4%,and the inter-assay average coefficient of variation was 9.7% and 11.5%,respectively.The detection sensitivity was 1∶ 1280.There was no cross-reactivity with classical swine fever virus（CSFV） and porcine parvovirus（PPV）.The stability test showed a relative deviation less than 25%.Conclusion The established ELISA method has good reproductivity,stability,specificity and sensitivity,and can be used in detection of pig JEV antibody.