ODA－H_2O_2－HBsAg－HRP伏安酶联免疫分析测定人血清乙型肝炎表面抗体的研究

Determination of HBsAb in Serum by Voltammetric Enzyme Immunoassay with Coupled Reaction

提出了一种偶合反应伏安酶联免疫分析测定人血清乙型肝炎表面抗体（ＨＢｓＡｂ）的新方法。该法将ＨＢｓＡｇ－ＨＲＰ催化Ｈ２Ｏ２氧化邻联茴香胺（ＯＤＡ）的反应与邻联茴香胺的氧化产物在电极上的反应相偶合，在ＢＲ缓冲溶液中，在－０．５６Ｖ（ＳＣＥ）左右产生灵敏的极谐波，根据测定标记在乙型肝炎表面抗原上的ＨＲＰ的量，求得发生免疫反应的乙型肝炎表面抗体的含量。本法对ＨＢｓＡｇ－ＨＲＰ及ＨＢｓＡｂ测定的灵敏度均高于经典的ＥＬＩＳＡ显色光度法。本法对ＨＢｓＡｂ的测定采用双抗原夹心法，用所建立的方法对实际病人血清样品进行了测定，并与现行的ＥＬＩＳＡ显色光度法进行对照，两种方法相关性很好。

A new voltammetric enzyme immunoassay with coupled reaction to determine HBsAbin human serum is develops. It is based on coupling the reaction ODA-H2O2 catalyzed by HBsAg-HRPwith the electro-reduction reaction of the oxidizing product of ODA to measure the activity of ABsAgHRP and the latter reaction exhibits a sensitive second order derivative linear sweep voltammetric waveat -0. 56V(SCE) in BR buffer,and so HBsAb will be measured.The sensitivity of the developed method to deteCt Hang-HRP is raised 10-fold when comparingwith the ELISA absorption metric method. The double antigen sandwish immunological method is usedto determine HBsAb. The linear relationship for diluted concentration of HBsAb positive contact serumis over 1: 1 ￣ 1: 512, and the sensitivity is improved 4-fold against the ELISA absorption metricmethod ; and the current ELISA absorption metric method is taken as a control method for those humanserum samples and the same results are obtained.