利用H9亚型的HA1蛋白建立ELISA抗体检测方法

摘要

The optimal working conditions of this ELISA method was investigated and determined:the recombi-nant protein diluted by 1:200, coating in the condition of 4℃overnight;the serum dilution was 1:50 and working concentration of HRP-labeled rabbit anti-protein IgG was 1:1000, both the incubation steps above were done at 37℃for 30min. The samples with optical dense(OD) greater than 0.109 were judged as cut off.%　　该试验对ELISA反应条件进行摸索，并确定了最适工作条件。结果表明，重组蛋白最适包被浓度为200倍稀释，最适包被条件为4℃过夜，血清稀释度为1颐50，HRP-兔抗鸡IgG的最适工作浓度为1颐1000，待检血清和酶标二抗的反应时间均37℃30min，底物在室温显色10min。根据已经建立的ELISA方法及反应条件，确定阴阳性临界值为0.109。