HIV-p24抗原的电化学免疫传感器研究

制备了一种基于巯基丁二胺铜(Ⅱ)(CuL)的自组装修饰传感器,用于人血清中HIV-p24水平免疫分析。其主要参数为:电极表面直径2 mm、高2 cm,CuL覆盖率为1.02×10-11mol/cm2,检测用血量<10 mL,检测速度<10 s。其电化学行为表现出表面控制过程。结合电化学酶联免疫分析法,在含有1.0 mmol/L H2O2的pH=6.2磷酸盐缓冲溶液中,获得HIV-p24的测定校正曲线为0.5~200 ng/mL,检测限为0.2 ng/mL。该传感器准确性、精确性和制备重复性良好。测定10 ng/mL HIV-p24时的变异系数为3.5%。该方法避免了常规电化学免疫测定中电子媒介体的加入,简化了分析步骤,缩短了分析时间。对艾滋病的诊断和检测具有一定价值。该研究的新颖性,已为宁波市科学技术情报研究所2006年9月20日出具的第20063360800348号《科技查新报告》所证实,属未见文献报道。

Electrochemical Sensor for Immunoassay of HIV-p24 Antigen

A sensor based on mercapto-butadiamine copper(CuL) monolayer modified gold electrode for determination of HIV-p24 antigen in human serum is proposed.The main parameters of the sensor are as follows.Diameter of the electrode surface is 2 mm and the height is 2 cm.CuL is self-assembled on Au electrode which gives a surface coverage of 1.02×10-11 mol/cm2.The electrode can detect HIV-p24 antigen within 10 s for blood volume less than 10 mL.The electrochemistry of the immobilized CuL displays a surface-controlled electrode process.Based on an electrochemical enzyme-linked immunoassay,the calibration curve was obtained ranging from 0.5 to 200 ng/mL and the detection limit is 0.2 ng/mL for HIV-p24 determination in pH=6.2 phosphate buffer solution containing 8 mmol/L H2O2.The sensor shows good accuracy,precision and reproducibility with the coefficient of variation of 3.5% at 10 ng/mL HIV-p24 concentration.This method avoids the addition of electron transfer mediator to the solution.The proposed method would be valuable for the diagnosis of AIDS disease.