Electrochemical studies of o-tilidine as a hydrogen donor substrate for peroxidase and its application in enzyme immunoassay

被引:14
作者
Jiao, K [1 ]
Sun, W [1 ]
机构
[1] Oingdao Inst Chem Technol, Dept Appl Chem, Shandong 266042, Peoples R China
基金
中国国家自然科学基金;
关键词
o-tilidine; horseradish peroxidase; enzyme immunoassay; cucumber mosaic virus; voltammetry;
D O I
10.1016/S0026-265X(01)00116-3
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
A highly sensitive electrochemical method is described for assay of horseradish peroxidase (HRP) using o-tilidine as substrate. Under the optimal conditions, the detection limit for HRP is 2.5 mU/l and the calibration range is from 5.0 to 1000 mU/l. The relative standard deviation of 11 measurements is 6.3% for 10.0 mU/l HRP. This new electrochemical system was further combined with an indirect enzyme immunoassay using direct antigen-coating format for the detection of cucumber mosaic virus (CMV). The results show an improved sensitivity over the traditional o-phenylenediamine (OPD) spectrophotometric enzyme-linked immunosorbent assay (ELISA) method. Using this technique, a minimum detectable level of 2.0 ng/ml of purified CMV and 1:12500 dilution of CMV in infected leaf extractions can be achieved. (C) 2002 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:123 / 130
页数:8
相关论文
共 23 条
  • [1] Flow injection electrochemical enzyme immunoassay based on the use of gold bands
    Abad-Villar, EM
    Fernández-Abedul, MT
    Costa-García, A
    [J]. ANALYTICA CHIMICA ACTA, 2000, 409 (1-2) : 149 - 158
  • [2] IMPROVED COLOR REAGENT FOR DETERMINATION OF BLOOD-GLUCOSE BY OXIDASE SYSTEM
    BARHAM, D
    TRINDER, P
    [J]. ANALYST, 1972, 97 (1151) : 142 - &
  • [3] ENZYME-LINKED IMMUNOADSORBENT ASSAY WITH ELECTROCHEMICAL DETECTION FOR ALPHA-1-ACID GLYCOPROTEIN
    DOYLE, MJ
    HALSALL, HB
    HEINEMAN, WR
    [J]. ANALYTICAL CHEMISTRY, 1984, 56 (13) : 2355 - 2360
  • [4] FOSSATI P, 1980, CLIN CHEM, V26, P227
  • [5] HE YN, 1997, TALANTA, V44, P923
  • [6] INRITSKII DM, 1992, ANAL CHIM ACTA, V261, P45
  • [7] Jiao K, 1997, ACTA CHIM SINICA, V55, P1121
  • [8] Application of p-phenylenediamine as an electrochemical substrate in peroxidase-mediated voltammetric enzyme immunoassay
    Jiao, K
    Sun, W
    Zhang, SS
    Sun, G
    [J]. ANALYTICA CHIMICA ACTA, 2000, 413 (1-2) : 71 - 78
  • [9] LOMMEL SA, 1982, PHYTOPATHOLOGY, V72, P1018, DOI 10.1094/Phyto-72-1018
  • [10] MONGET D, 1978, AM BIOL CLIN, V6, P527