高等学校化学学报

高等学校化学学报 ›› 2000, Vol. 21 ›› Issue (8): 1200.

• 论文 • 上一篇    下一篇

MAP-H2O2-HRP伏安酶联免疫分析测定南方菜豆花叶病毒

张书圣1, 焦奎1, 陈洪渊2, 王曼霞1, 张成良3, 张作芳3   

  1. 1. 青岛化工学院应用化学系, 青岛 266042;
    2. 南京大学化学系, 南京 210093;
    3. 农业部植物检疫实验所, 北京 100029
  • 收稿日期:1999-11-17 出版日期:2000-08-24 发布日期:2000-08-24
  • 通讯作者: 焦　奎(1945年出生),男,教授,博士生导师,从事电分析化学研究.
  • 基金资助:

    国家自然科学基金(批准号:29775012);山东省青年科学基金(批准号:Q99B16)

MAP-H2O2-HRP Voltammetric Enzyme linked Immunoassay for Trace Southern Bean Mosaic Virus

ZHANG Shu-Sheng1, JIAO Kui1, CHEN Hong-Yuan2, WANG Man-Xia1, ZHANG Cheng-Liang3, ZHANG Zuo-Fang3   

  1. 1. Departmentof App lied Chemistry, Qingdao Instituteof Chemical Technology, Qingdao 266042, China;
    2. Departmentof Chemistry, Nanjing University, Nanjing 210093, China;
    3. Instituteof Plant Quarantine, Ministry of Agriculture, Beijing 100029, China
  • Received:1999-11-17 Online:2000-08-24 Published:2000-08-24

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摘要： 提出间氨基酚(MAP)-H2O2-辣根过氧化物酶(HRP)伏安酶联免疫分析新体系,并用于南方菜豆花叶病毒(SBMV)的测定.以线性扫描二阶导数伏安法检测HRP催化H2O2氧化MAP的产物,用于游离HRP及SBMV的测定,灵敏度均高于经典的ELISA显色光度法.本法对HRP测定的线性范围为1.0×10-8～1.0×10-6g/L,检测限为3.8×10-9g/L;对SBMV测定的线性范围为4.0～5000ng/mL,检测限为4.0ng/mL.用所建立的方法测定病毒感染病叶澄清液的最高稀释比为1∶1.5×105,并与现行的ELISA显色光度法进行对照,二者相关性很好.

关键词: 南方菜豆花叶病毒, 辣根过氧化物酶, 间氨基酚, 免疫分析, 伏安法

Abstract: Avoltammetric enzyme-linked immunoassay based on the new system of m-aminophenol(MAP)-H2O2-horseradish peroxidase(HRP) has been developed and used for the detection of HRPand southern bean mosaic virus(SBMV). HRPor labelled HRPcatalyzes the oxidation reaction of MAPwith H2O2, the product of which produces a sensitive voltammetric peak at potential of -0.46V(vs. SCE ) in Britton-Robinson(B-R) buffer solution.By using this voltammetric peak, HRPcan be measured with a detection limit of 38×10-9 g/Land a linear range of 10×10-8 —10×10-6 g/L. The detection limit for the clarified SBMVis 4.0 ng/mLand the linear range of 4.0—5000 ng/mL. The highest dilution ratio detected for the infected leaf sap is 1∶1.5×105.

Key words: Southern bean mosaic virus(SBMV), Horseradish peroxidase(HRP), m-Aminophenol, Immunoassay, Voltammetry

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