MAP-H_2O_2-HRP伏安酶联免疫分析测定南方菜豆花叶病毒

MAP-H_2O_2-HRP Voltammetric Enzyme linked Immunoassay for Trace Southern Bean Mosaic Virus

提出间氨基酚 ( MAP) -H2 O2 -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 H2 O2 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× 1 0 - 8～1 .0× 1 0 - 6 g/L,检测限为 3 .8× 1 0 - 9g/L;对 SBMV测定的线性范围为 4 .0～ 50 0 0 ng/m L,检测限为 4 .0ng/m L.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× 1 0 5 ,并与现行的 ELISA显色光度法进行对照 。

A voltammetric enzyme linked immunoassay based on the new system of m aminophenol(MAP) H 2O 2 horseradish peroxidase(HRP) has been developed and used for the detection of HRP and southern bean mosaic virus(SBMV). HRP or labelled HRP catalyzes the oxidation reaction of MAP with H 2O 2, the product of which produces a sensitive voltammetric peak at potential of -0 46 V(vs. SCE ) in Britton Robinson(B R) buffer solution.By using this voltammetric peak, HRP can be measured with a detection limit of 3 8×10 -9 g/L and a linear range of 1 0×10 -8 —1 0×10 -6 g/L. The detection limit for the clarified SBMV is 4 0 ng/mL and the linear range of 4 0—5 000 ng/mL. The highest dilution ratio detected for the infected leaf sap is 1∶1 5×10 5.