HIV-p24抗原的电化学免疫传感器研究

Electrochemical Sensor for Immunoassay of HIV-p24 Antigen

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2mm、高2cm，CuL覆盖率为1．02×10^-11 mol/cm^2，检测用血量〈10mL，检测速度〈10S。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1．0mmol/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0．5-200ng/mL，检测限为0．2ng/mL。该传感器准确性、精确性和制备重复性良好。测定10ng/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。

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^-11mol/cm^2. 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 8mmol/L H2O2. The sensor shows good accuracy, precision and reproducibility with the coefficient of variation of 3.5% at 10ng/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.