氯化血红素作为模拟酶荧光免疫分析乙肝表面抗原

作     者：朱庆枝 郑雪英 李芳 许金钩 刘风华 李文友 

作者机构：国家教育委员会材料和生命过程分析科学开放实验室厦门361005 厦门大学化学系厦门大学生物系厦门361005 

出 版 物：《高等学校化学学报》 (Chemical Journal of Chinese Universities)

年 卷 期：1997年第18卷第8期

页      面：1303-1305页

核心收录：

学科分类：1004[医学-公共卫生与预防医学(可授医学、理学学位)] 1010[医学-医学技术（可授医学、理学学位）] 100401[医学-流行病与卫生统计学] 10[医学] 

基　　金：国家教育委员会材料和生命过程分析科学开放实验室资助 

主　　题：模拟酶 氯化血红素 荧光免疫分析 HBsAg 维生素B 

摘      要：A novel mimetic enzyme immunoassay method was developed for the determina-tion of hepatitis B surface antigen (HBsAg) in solution. Hemin, a horseradish peroxidase substitute, was used as a labelling reagent to catalyze the reaction of thiamine, a new fluoro-genic substrate, with hydrogen peroxide at pH8.5. In the sandwich immunoassay,anti-HBsAg antibody was coated on a 96-well plate(polystyrene), which first reacted with stan-dard HBsAg solution, and then further reacted with the fixed amount of hemin-labelled anti-HBsAg. After the two-step immunoreaction, the immunochemically adsorbed hemin-anti-HBsAg conjugate moiety was determined by measuring the fluorescence produced in a solu-tion containing thiamine and hydrogen peroxide. The flourescence intensity was directly pro-portional to the concentration of HBsAg. The calibration graph for HBsAg was linear over the range 2.5~500ng/well with a detection limit of 2.5ng/well.