胶体金免疫层析技术同步检测甲、戊型肝炎病毒IgM抗体的研究

The study of colloidal gold immunochromatography assay for the synchronous detection of hepatitis A and E virus-specific IgM antibody

目的 建立胶体金免疫层析 (GICA)同步检测血清中的甲、戊型肝炎病毒的IgM抗体的方法。方法 采用胶体金标记鼠抗人IgM单克隆抗体 ,制成免疫层析测试条。血清中甲、戊型肝炎的特异性抗体与测试条上金标抗体结合后沿着硝酸纤维素膜移动与膜上固相包被的抗原 (HAAg、HEAg)、抗体 (羊抗鼠IgM)结合形成肉眼可见的红色线条。结果 自制的GICA试条与EIA对比检测血清标本 188份 ,各单项指标IgM 抗HAV、IgM 抗HEV两法总符合率依次为97.9%、98.4 % ,检测灵敏度可达 2ng/ml。结论 用于同步检测血清中的甲、戊型肝炎病毒特异性抗体的GICA试条的制备条件已基本建立 ;其检测结果特异性强、灵敏度高、操作方法简便快捷且无需特殊仪器设备。

Objective To develop gold immunochromatography assay (GICA) for the synchronous detection of hepatitis A and E virus-specific immunoglobulin M(IgM) antibody in sera.Methods Colloidal golds were couple with monoclonal antibody (MAH IgM) to prepare an GICA apparatus . The gold antibodies conjugate reacts with hepatitis A and E virus-specific antibodies in serum and migrates up the nitrocellose strip on which hepatitis A antigen and hepatitis E antigen were immobilized , thus producing the detection reaction . Results 188 sera of patients and healthy people were detected by GICA and EIA .The corresponding rate of both tests for anti-HAV-IgM was 96.8% and for anti-HEV-IgM was 97.5% ,The sensitivity of GICA was 2ng/ml by detecting Anti-HAV IgM or Anti-HEV IgM panel from the National Institute for the control of Pharmaceutical and Biological Produts.Conclusion The hepatitis A and E virus-specific antibodies in serum can be simultaneously detected in one-step. GICA is a sensive , specific , simple and rapid assay.