纳米磁性微球免疫伏安法测定乙肝表面抗原  

Detection of Hepatitis B Surface Antigen Using Immunovoltammetry with Nano-Magnetic Microsphere

作　　者：程琼[1,2] 彭图治[1] 刘爱丽[1] 

机构地区：[1]浙江大学化学系,西溪校区 [2]嘉兴学院生物与化工学院,嘉兴314001

出　　处：《化学学报》2004年第24期2447-2450,共4页Acta Chimica Sinica

基　　金：国家自然科学基金 (Nos.2 99750 2 4 ;2 0 2 750 34);浙江省重点科技计划 (No.2 0 0 3C2 1 0 2 4 )资助项目

摘　　要：采用化学键合法将乙肝抗体固化在自行制备的纳米磁性高分子功能微球表面 ,利用免疫夹心反应原理 ,捕获溶液中的乙肝表面抗原和标记有辣根过氧化物酶的乙肝第二抗体 .在外加磁场的作用下 ,抗体抗原结合物从样品溶液中分离 ,在含有邻氨基苯酚和过氧化氢的底液中 ,生成具有电活性的化合物 3 氨基吩呃嗪 ,用示差脉冲伏安法进行测定 .响应电流与乙肝表面抗原浓度分别在 0 2～ 1 0和 1 0～ 5 0 0ng·mL-1成线性关系 ,检出限达 0 0 60ng·mL-1.采用本方法检测血清中乙肝表面抗原 ,灵敏度大大高于目前临床采用的酶联免疫吸附法 .A novel method for detecting hepatitis B surface antigen (HBsAg) using nano-magnetic polymer carboxyl microsphere was reported. The surface of the nano-magnetic polymer carboxyl microsphere covalently bound with hepatitis B surface antibody (HBsAb), and then captured HBsAg and hepatitis B secondary antibody labeled with horseradish peroxide enzyme. The microsphere with sandwich complex was separated magnetically from sample solution and the enzyme labeled at the complex catalyzed the reaction of 2-aminohydroxybenzene and hydrogen peroxide in the buffer solution. The electroactive product 2-hydroxy-3-aminophenoxazine was detected using differential pulse voltammetry. The peak current was linear with the concentration of HBsAg in the range of 0.2 - 1.0 and 1.0 similar to 500 ng . mL respectively. The detection limit was found to be 0.06 ng . mL(-1). The method has been used to detect the concentration of hepatitis B virus in human serum and the sensitivity was much higher than that of enzyme-linked immunosorbent assay (ELISA) used in diagnostic analysis.

关 键 词：乙肝表面抗原 乙肝抗体 酶联免疫吸附法 样品溶液 磁性微球 临床 血清 邻氨基苯酚 纳米 反应原理 

分 类 号：O657.1[理学—分析化学]