流行性乙型脑炎病毒IgG抗体捕获酶联免疫吸附试验检测方法的建立
Establishment of IgG capturing enzyme-linked immunosorbant assay to detect Japanese encephalitis virus antibody
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摘要: 目的 利用流行性乙型脑炎(乙脑)病毒特异性单克隆抗体建立检测人血清中乙脑病毒IgG抗体的捕获ELISA方法。 方法 将乙脑病毒特异性单克隆抗体包被在固相载体上,加入灭活并经过纯化的乙脑病毒抗原,再加入待测血清,经洗涤、显色等判定最终检测结果。采用免疫荧光法作为对照。 结果 与免疫荧光法相比较,本方法的灵敏度为95.6%,特异度为85.7%,一致性为91.3%;本方法的灵敏度和特异度高于市售的两种检测试剂。 结论 本研究所建立的乙脑病毒IgG抗体捕获ELISA方法具有良好的检测性能,可以应用于正常人群血清中乙脑病毒IgG抗体的筛查。Abstract: Objective To establish IgG capturing enzyme-linked immunosorbant assay (ELISA) to detect Japanese encephalitis virus (JEV) antibody in human serum by using monoclonal antibody. Methods Monoclonal antibody to JEV was coated in the plate, inactivated and purified JEV antigen was added to the coated plate and then detected serum was added. The results were interpreted according to color development. Immunofluorescence assay (IFA) was used as the control method in this study. Results Compared with IFA, the sensitivity, specificity and consistency of the developed ELISA was 95.6%, 85.7% and 91.3% respectively. Its sensitivity and specificity was higher than other two commercial kits. Conclusion The developed ELISA has good proficiency and can be used to screen IgG antibody to JEV in human serum.
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