定量检测人狂犬病毒中和抗体间接酶联免疫吸附测定方法的建立

Development of an indirect ELISA for quantitative detection of neutralizing antibodies against human rabies

目的建立定量检测人狂犬病毒中和抗体间接酶联免疫吸附测定(ELISA)方法。方法选择包被抗原和酶标二抗的最适浓度,用人狂犬病毒抗体标准品标化系列工作标准品,对临床收集的各类血清标本进行检测和分析。结果间接ELISA定量检测中和抗体的最佳包被糖蛋白抗原浓度为10μg/L,酶标二抗的工作浓度为1∶4 000,临床检验,间接ELISA定量方法灵敏度为100%,特异度为98.2%,总符合率99.0%;与快速荧光灶抑制实验呈(RFFIT)高度相关性(r=0.981)。结论本研究建立的定量检测狂犬病中和抗体方法,可以用于人狂犬病毒中和抗体的定量检测。

Objective To develop an indirect enzyme linked immunosorbent assay （ELISA） for quantitative detection of neutralizing antibodies against human rabies. Methods The optimal working concentrations of coating antigen and HRP-labeled mice anti bovine IgG were determined using antibodies against human rabies as the working standard, and serum specimen were detected and analyzed. Results The optimum test conditions were as follows：the concentration of neutralizing antigen was 10 μg/L, HRP labeled mice anti bovine IgG was 1 ： 4 000. The sensitivity, specificity, coincidence rate of indirect EI.ISA method in clinical laboratory test were 100%, 98.2%, 99.0%, respectively,and the correlation coefficient between indirect ELISA method and RFFIT was 0. 981. Conclusion The results indicated that the indirect ELISA could be used for detection of neutralizing antibodies against human rabies.