基于HRP-H_2O_2-OPDA发光体系的流感病毒H1N1酶联免疫传感器

Study on an enzyme linked immunsensor for influenza virus H1N1 based on HRP-H_2O_2-OPDA fluorescence system

制备了一种基于HRP-H_2O_2-OPDA发光体系的流感病毒H1N1酶联免疫传感器。利用流感病毒H1N1表面的HA(血球凝集素)蛋白与糖蛋白能有效结合的特性,选择辣根过氧化物酶(HRP)作为生物标记物直接标记在流感病毒的表面,HRP能有效催化H_2O_2-邻苯二胺(OPDA)体系产生灵敏的显色反应和荧光效应。实验对HRP孵化时间、H_2O_2-OPDA催化反应时间等参数进行了优化。结果表明,该酶联免疫传感器对流感病毒H1N1检测的动态响应范围为0.0001~0.5μg/m L,检出限为50 pg/m L(3σ)。其他亚型病毒和干扰蛋白对该传感器的干扰较小,表现出良好的选择性。所构建的方法可为其它流感病毒及其它抗原的检测提供参考。

An enzyme linked immunsensor for influenza virus H1N1 recognition based on HRP–H2O2–OPDA fluorescence system was prepared. Taking advantage of the combination characteristics between hemagglutinin（ HA） on the surface of influenza virus H1N1 and horseradish peroxidase（ HRP）,which is a type of active glycoprotein,HRP,as a biomarker was labeled directly on the surface of virus. HRP can effectively catalyze the H2O2– OPDA（ o-phenylenediamine） to produce significant color reaction and sensitive fluorescent effect. The effects of HRP incubation time,H2O2–OPDA catalytic reaction time on the performance of the assay were studied and optimized. Results showed that the dynamic response range between the enzyme linked immunsensor and the concentration of influenza virus H1N1 over the range of 0. 0001 - 0. 5 μg/m L with a limit of detection（ LOD,3σ）.At the same time,the sensor with good selectivity has less interference to other subtype of influenza virus H5N1 and interfering protein. The constructed sensor possessed a great deal of properties,such as convenient,high sensitivity,high selectivity,also provided a promising potential for recognizing other subtypes of influenza virus and other antigens.