联苯胺-H_2O_2-HRP伏安酶联免疫分析体系测定植物病毒TMV和TRSV  

Determination of TMV and TRSV by Voltammetric Enzyme \=Linked Immunoassay with BenzidineH2O2HRP System

作　　者：牛淑妍[1,2] 焦奎[1,2] 张成良 张作芳[1,2] 刘靖宇 魏澎[3] 

机构地区：[1]青岛化工学院应化系 [2]农业部植物检疫实验所 [3]核工业部北京地质研究所

出　　处：《分析科学学报》1999年第3期195-198,共4页Journal of Analytical Science

基　　金：国家自然科学基金;山东省自然科学基金

摘　　要：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 voltammetric enzymelinked immunoassay with BenzidineH2O2HRP system for the determination of Tobacco Mosaic Virus (TMV), and Tobacco Ringspot Virus(TRSV) is developed. It is based on coupling the reaction of BenzidineH2O2 catalyzed by IgGHRP with the electroreduction reaction of the oxidizing product of Benzidine to measure the activity of the immunocomplex TMVAg:TMVAs:IgGHRP (or TRSVAg:TRSVAs:IgGHRP ) and the latter reaction exhibits a sensitive second order derivative linear sweep voltammetric peak at -0.62 V (vs.SCE) in BR buffer solution . Therefore, TMV and TRSV can be measured sensibly. The sensitivity of the developed method for the detection of TMV and TRSV is higher than ELISA spectrophotometric method.\=

关 键 词：酶联免疫分析 烟草花叶病毒 烟草环斑病毒 测定 

分 类 号：S435.72[农业科学—农业昆虫与害虫防治]