甲型肝炎病毒IgM AlphaLISA检测方法的初步建立

Primary establishment of an alphaLISA assay for detection of HAV IgM

目的建立甲型肝炎病毒抗体IgM的AlphaLISA检测。方法浓缩甲型肝炎病毒抗原并生物素化，通过dot—Blot法摸索出生物素化的甲型肝炎病毒的最适量为6×10-8ng。优化供体珠、受体微珠、血清等实验反应条件，建立了甲型肝炎病毒的AlphaLISA的IgM检测方法。并对23份感染甲型肝炎病毒的IgM阳性患者和70份健康人的血清进行检测。结果AlphaLISA甲型肝炎病毒的IgM方法检测：灵敏度（真阳性率）为78％；特异度（真阴性率）为98．5％；假阳性率为1％；假阴性率为22％；阳性预测值为95％；阴性预测值为93％；准确度为94％。结论建立了均相、快速检测AlphaLISA甲型肝炎病毒IgM检测方法并得到初步应用。

Objective To develop an AlphaLISA method for detection of antibody of Hepatitis A virus. Methods After hepatitis A virus antigen was concentrated and biotinylated, optimal biotinylated HAV antigen, donor bead, aceeptor bead concentration have been explored and determined by both dot- blotting and AlphaLISA methods. 97 samples including 23 serums from patients HAV infected and 70 serums from blood donors have been detected by new AIphaLISA method. Results The sensitivity and specificity for anti-HAV IgM were 78% and 98.5% , the positive and negative predictive value were 95% and 93%. Conclusion A homogeneous and fast AlphaLISA assay for detection of HAV IgM has been established by preliminary verification on samples.