戊型肝炎病毒IgM抗体检测试剂盒制备及功能验证

Preparation and Functional Validation of Hepatitis E Virus IgM Antibody Detection Kit

目的:建立一种快速、简单实用的检测戊型肝炎病毒IgM抗体的胶体金免疫层析法。方法:用柠檬酸三钠还原法制备胶体金颗粒,用鼠源人IgM单克隆抗体进行胶体金标记,采用胶体金免疫层析技术及间接法原理检测戊型肝炎病毒IgM抗体,优化抗体最适宜浓度、抗体稀释度及包被抗原浓度,利用市售ELISA免疫吸附试剂对100例临床血清标本进行比较实验。结果:鼠抗人-IgM抗体标记最适合蛋白用量为10μg/mL、金标鼠抗人-IgM抗体最适合的稀释度为8%、最佳抗原包被浓度为1.0 mg/mL;与酶联分析法对比,阳性符合率98.1%,阴性符合率100%。结论:HEV-IgM胶体金免疫层析试剂,方法简便、快速,适应戊型肝炎早期感染的快速检测及科研工作需要。

Objective:To establish a rapid,simple and practical colloidal gold immunochromatographic method for the detection of hepatitis E virus IgM antibody.Methods:Colloidal gold particles are prepared by trisodium citrate reduction method,colloidal gold labeled with murine human IgM monoclonal antibody,hepatitis E virus IgM antibody is detected by colloidal gold immunochromatography technology and indirect method,the optimal concentration of antibody,antibody dilution and coating antigen concentration are optimized,and 100 clinical serum specimens are compared by commercially available ELISA immunosorbent reagent.Results:The most suitable protein dosage for murine anti-human-IgM antibody labeling is 10μg/mL,the most suitable dilution of gold standard mouse antihuman-IgM antibody is 8%,and the optimal antigen coating concentration is 1.0 mg/mL;compared with the enzymelinked analysis,the positive coincidence rate is 98.1%and the negative coincidence rate is 100%.Conclusion:HEVIgM colloidal gold immunochromatographic reagent is a simple and rapid method,which is suitable for the rapid detection and scientific research of early hepatitis E infection.