动物狂犬病中和性抗体竞争ELISA检测试剂盒的研制

目的研制一种能够定量检测狂犬病血清中和抗体的直接竞争ELISA试剂盒,以满足我国动物狂犬病免疫监测的需要。方法以表达狂犬病病毒糖蛋白的真核细胞培养物为包被抗原,捕获抗体为具有病毒中和活性的酶标单克隆抗体,标准血清为经荧光抗体病毒中和试验（FAVN）准确定量的犬血清。结果本试剂盒的最低检测限为0.25U/ml,线性检测范围为0.25～8U/ml,标准曲线的平均板内变异系数为2.8%,板间平均变异系数为4.9%,在4℃下至少可以存放6个月。结论本研究提供了一种操作安全、定量准确的动物狂犬病中和抗体检测试剂盒,具有良好的实际应用前景。

Development of a direct competitive ELISA kit for monitoring the level of rabies neutralizing antibody in animals

Objective To develop a direct competitive ELISA kit for quantitative detection of rabies neutralizing antibody in serum in order to fulfill the needs of monitoring animal rabies immunity.Methods The kit was based on monoclonal antibody against rabies glycoprotein for the measurement of rabies virus-specific neutralizing antibodies from serum samples of animals.Data generated by this ELISA test indicated a strong correlation for accuracy when compared with the gold standard fluorescent antibody virus neutralization（FANV）.Results The linear range of detection by the kit was from 0.25 to 8 U/ml,with minimum detection limit being 0.25 U/ml and the intra-assay and inter-assay standard deviation being 2.8% and 4.9%,respectively.Conclusion The major advantages of the c-ELISA kit are that it is a quantitative detection method that can be completed in 2 hours,requires no use of live virus and can be performed without specialized laboratory apparatus.