禽流感病毒H9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 H9 subtype avian influenza virus

摘要: 采用禽流感病毒多克隆抗体及H9亚型特异性单克隆抗体,研究建立H9亚型特异性抗原捕捉ELISA检测方法,用于检测H9亚型禽流感病毒.优化了反应条件,确定了包被抗体、检测抗体及酶结合物的最佳工作浓度,对该方法的敏感性、特异性、重复性及稳定性分析,并与RT-PCR方法比较.通过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H9亚型禽流感病毒.

Abstract: By using avian influenza polyclonal antibody and H9 subtype-specific monoclonal antibody,a H9 subtype specific antigen capture ELISA was developed for detection of H9 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 compared with RT-PCR.The results indicated that antigen capture ELISA was sensitive,specific,with high repeatability and stability for detection of H9 subtype avian influenza virus in clinical samples,embryo and cell culture tissues.