禽流感病毒H5亚型特异性抗原捕捉ELISA检测方法的研究

Development of subtype-specific antigen capture ELISA for detection of H5 subtype avian influenza virus

在获得禽流感病毒多克隆抗体及H5亚型特异性单克隆抗体的基础上,研究建立H5亚型特异性抗原捕捉ELISA检测方法,用于检测H5亚型禽流感病毒.优化反应条件,确定包被抗体、检测抗体及酶结合物的最佳工作浓度,进行敏感性、特异性、重复性及稳定性分析,并与RT-PCR方法比较.同时使用该方法对野外样品进行检测.结果表明:该方法敏感、特异,具有良好的重复性和稳定性,可用于检测临床样品、鸡胚培养物及细胞培养物中的H5亚型禽流感病毒.

Based on obtaining avian influenza polyclonal antibody and H5 subtype-specific monoclonal antibody, a H5 subtype specific antigen capture ELISA was developed for detection of H5 subtype avian influenza virus. The reactive condition and the working titers of coating antibody, detective antibody and enzyme-labeled conjugate were optimized,and the assay sensitivity, specificity, repeatability, stability were analyzed and com- pared with virus isolation and identification. The results indicated that antigen capture ELISA was sensitive, specific, with high repeatability and stability for detection of H5 subtype avian influenza virus in clinical sampies, embryo and cell culture tissues.