无标记检测CD4^+T淋巴细胞的免疫传感器构建  

Immunosensor construction of label free detection of CD4^+T lymphocytes

作　　者：蒋兴宇[1] 蒋栋能[1] 孟凡飞[1] 蒲晓允[1] 

机构地区：[1]第三军医大学新桥医院检验科,重庆400037

出　　处：《国际检验医学杂志》2017年第6期753-755,759,共4页International Journal of Laboratory Medicine

基　　金：国家自然科学基金资助项目(81472034)

摘　　要：目的构建一种无标记检测CD4^+T淋巴细胞的免疫传感器。方法采用葡萄球菌A蛋白(SPA)法定向固定单克隆抗体于金叉指微阵列电极表面以捕获CD4^+T淋巴细胞,并用循环伏安法(CV)扫描对金叉指微阵列电极表面的修饰情况进行表征,最后通过电化学交流阻抗谱法(EIS)对免疫传感器捕获的CD4^+T淋巴细胞进行阻抗检测,通过等效电路拟合获得的阻抗值变化绘制标准曲线。结果该免疫传感器检测CD4^+T淋巴细胞的线性范围是(5.0×103-5.0×106)/mL,检测下限为5.0×102/mL。结论该免疫传感器的结果准确可靠,检测过程简便,价格低廉,有望用于实时检测系统,为实现快速、准确、价廉的CD4^+T淋巴细胞分类计数提供帮助。Objective To construct an immunosensor for detecting CD4^＋T lymphocytes without labeling.Methods The staphylococcus protein A（SPA）method was adopted to conduct the oriented immobilization of CD4 monoclonal antibodies on the gold interdigitated microelectrode surface for capturing CD4^＋T lymphocytes.Then cyclic voltammetry（CV）method was used to conduct the representation of modification situation on the gold interdigitated microelectrode surface.Finally the electrochemical impedance spectroscopy（EIS）was used to detect the impedance of CD4^＋T lymphocytes captured by the immunosensor.The standard curve was drawn by the impedance values change obtained by the equivalent electric circuit fitting.Results The linear range of this immunosensor for detecting CD4^＋T lymphocytes was（5×10^3-5.0×10^6）/mL,with lower detection limit of 5.0× 10^2/mL.Conclusion The constructed immunosensor has accurate and reliable detection results uhidn is simple to operate accurate,convenient and cheap,which might be expected to be used in the real-time detection system,and offers help for realizing rapid,accurate and inexpensive CD4^＋T lymphocyte count.

关 键 词：叉指微阵列电极 免疫传感器 葡萄球菌A蛋白 交流阻抗谱 

分 类 号：R446.6[医药卫生—诊断学] TP212[医药卫生—临床医学]