邻苯二胺-过氧化氢-辣根过氧化物酶酶联免疫示差脉冲伏安法测定人血清类风湿因子抗体  

o -Phenylenediamine -Hydrogen Peroxide -Horseradish Peroxidase Differential pulse Vollammetric Immunoassay for Determination of Rheumatoid Factor in Human Serum

作　　者：何亚楠[1] 陈洪渊[1] 虞伟[2] 武建国[2] 

机构地区：[1]南京大学化学系,南京210093 [2]南京军区总医院,南京210000

出　　处：《分析化学》1997年第1期30-33,共4页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金资助课题。

摘　　要：建立了邻苯二胺（ＯＰＤ）－Ｈ２Ｏ２－辣根过氧化物酶（ＨＲＰ）酶联示差脉冲伏安分析体系并用于测定人血清中类风湿因子（ＲＦ），ＨＲＰ催化Ｈ２Ｏ２氧化ＯＰＤ所形成酶催化产物在ｐＨ２．０磷酸盐－枸橼酸缓冲溶液中于－０．１８ Ｖ左右产生一灵敏示差脉冲伏安峰，ＲＦ浓度在１．２５～２０．０ Ｕ／ｍＬ之间与峰电流呈线性关系，应用此峰检测人血清ＲＦ的检测限低至 ０．２８Ｕ／ｍＬ。该法较相同条件下ＥＬＩＳＡ显色光度测定法的灵敏度增加５倍，且受干扰较少。A differential pulse voltammetric immunoassay using o-phenylenediamine-hydro- gen peroxide-horseradish peroxidase (OPD-H2O2-HRP) for the determination of rheumatoid factor(RF) antibody was developed. The enzymatic reaction product produced by HRP cat- alyzed oxidation of OPD with H2O2 exhibits a sensitive voltammetric response at - 0. l8 V (vs. Ag/AgCl) in pH 2. 0 phosphate-citrate buffer solution. The peak current is linear with RF concentration in the range of 1. 25-20. 0 U/mL. The detection limit for RF antibody is 0. 28 U/mL, which was five times lower than that obtained by traditional spectrophotometric enzyme-linked immunoassay procedure. The proposed method has been applied to the deter- mination of RF in human serum with satisfactory results.

关 键 词：类风湿性关节炎 类风湿因子 示差脉冲伏安法 

分 类 号：R593.210.4[医药卫生—内科学]