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提出了联苯胺-H2O2-辣根过氧化物酶(HRP)伏安酶联免疫分析体系测定植物病毒烟草花叶病毒(TMV)和烟草环斑病毒(TRSV)的新方法。HRP标记的羊抗兔酶标记抗体IgG-HRP可以催化H2O2氧化联苯胺的反应,其氧化产物在Briton-Robinson(BR)缓冲溶液中在-0.62V(SCE)左右产生灵敏的线性扫描二阶导数伏安峰,可以测定IgG-HRP。根据IgG-HRP与植物病毒及其抗血清的免疫反应,可以间接测定植物病毒。本法测定TMV的检出限为0.25ng/mL,线性范围为0.25~5000ng/mL;测定TRSV的检出限为1.5ng/mL,线性范围为1.5~3000ng/mL;测定TMV烟草病叶澄清液的最高稀释比为1∶10000。检测灵敏度高于酶联免疫吸附显色光度法(ELISA)
A new method for determination of TMV and TRSV by benzidine-H2O2-horseradish peroxidase (HRP) voltammetric enzyme-linked immunoassay was proposed. The HRP-labeled goat anti-rabbit antibody labeled IgG-HRP catalyzes the oxidation of benzidine by H2O2. The oxidation products produce sensitive second-order linear sweeps at -0.62 V (SCE) in a Briton-Robinson (BR) Derivative voltammetric peak, IgG-HRP can be determined. Based on the immune response of IgG-HRP to plant viruses and their antisera, plant viruses can be indirectly measured. The detection limit of TMV was 0.25ng / mL and the linear range was 0.25-5000ng / mL. The detection limit of TRSV was 1.5ng / mL and the linear range was 1.5-3000ng / mL. The maximum dilution of TMV tobacco leaf extract was 1: 10000. Detection sensitivity higher than enzyme-linked immunosorbent colorimetry (ELISA)