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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
引用本文:何亚楠,陈洪渊,虞伟,武建国.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[J].分析化学,1997(1).
作者姓名:何亚楠  陈洪渊  虞伟  武建国
作者单位:南京大学化学系!南京210093(何亚楠,陈洪渊),南京军区总医院!南京210000(虞伟,武建国)
基金项目: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。

关 键 词:类风湿因子  酶联免疫吸附法  示差脉冲伏安法  邻苯二胺

o -Phenylenediamine -Hydrogen Peroxide -Horseradish Peroxidase Differential pulse Vollammetric Immunoassay for Determination of Rheumatoid Factor in Human Serum
He Yanan,Chen Hongyuan.o -Phenylenediamine -Hydrogen Peroxide -Horseradish Peroxidase Differential pulse Vollammetric Immunoassay for Determination of Rheumatoid Factor in Human Serum[J].Chinese Journal of Analytical Chemistry,1997(1).
Authors:He Yanan  Chen Hongyuan
Abstract: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.
Keywords:Rheumatoid factor  differential pulse voltammetry  o-phenylenediamine
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