基于三重信号放大的电化学传感器检测黄体酮  

Detection of progesterone by electrochemical sensor based on triple signal amplification

作　　者：蔡杰 杨雅妮 罗大娟 高荣 陆发英 刘冰倩 CAI Jie;YANG Yani;LUO Dajuan;GAO Rong;LU Faying;LIU Bingqian(School of Pharmacy,Guizhou University,Synthetic Pharmaceutical Engineering Laboratory of Guizhou Province,Guiyang 550025)

机构地区：[1]贵州大学药学院,贵州省合成药物工程实验室,贵阳550025

出　　处：《分析试验室》2020年第9期1019-1023,共5页Chinese Journal of Analysis Laboratory

基　　金：国家自然科学基金(21864007);贵州省自然科学基金(黔科合平台人才[2018]5781;黔科合基础[2020]1Y042)项目资助。

摘　　要：基于DNA串联体(DNA concatamers)-G四聚体(G-quar DNA)-核酸外切酶Ⅰ (Exo Ⅰ)三重信号放大策略,以电沉积金的玻碳电极(dep Au/GCE)为反应平台,示差脉冲伏安法(DPV)为检测手段,构建简便灵敏的黄体酮(Pro)检测方法。基于碱基互补配对,富含鸟嘌呤(G)的辅助链1(H1)和辅助链2(H2)顺次与修饰在dep Au/GCE表面的目标DNA (t DNA)杂交形成DNA concatamers。氯化血红素(Hemin)与H1和H2未配对部分结合形成G-quar DNA分布于DNA concatamers四周。亚甲蓝(MB)稳定嵌入DNA concatamers和G-quar DNA中,增大电信号。当Pro存在时,Pro与t DNA特异性结合触发G-quar DNA/DNA concatamers从电极表面脱落。Exo Ⅰ特异性剪切Pro/t DNA结构,释放出的Pro进入下一轮循环,从而实现级联信号放大作用。电信号强度与Pro浓度呈负相关。峰电流与Pro质量浓度在1~40 ng/mL范围内呈良好的线性关系,检出限为0.1 ng/mL。Based on the triple signal amplification strategy of DNA concatamers-G-quadruplex DNA( G-quar DNA)-ExonucleaseⅠ( Exo Ⅰ),a simple and sensitive progesterone( Pro) detection method was constructed by means of differential pulse voltammetry( DPV) and glassy carbon electric reaction platform of electrodeposition gold( dep Au/GCE). Based on the base complementary pairing principle,auxiliary chains 1( H1) and 2( H2)rich in guanine( G) were successively hybridized and combined with the target DNA strand( tDNA) modified on the surface of dep Au/GCE to form the DNA concatamers. Hemin combined with unpaired H1 and H2 to form Gquadruplex structure,which was distributed around the DNA concatamers structure. The embedding of methylene blue( MB) enhanced the stability of the DNA concatamers and G-quadruplex structures,and improved the current signal values. In the presence of Pro,the t DNA-specific binding of the target triggered G-quar DNA/DNA concatamers to detach from the electrode surface. Exo Ⅰ specifically clipped the Pro/t DNA structure,releasing the Pro into the next cycle,thus achieving cascade signal amplification. The electrical signal strength is negatively correlated with Pro concentration. The peak current showed a good linear relationship with Pro concentration in the range of 1-40 ng/m L,with the detection limit of 0. 1 ng/m L.

关 键 词：黄体酮 核酸外切酶Ⅰ DNA串联体 G四聚体 信号放大 

分 类 号：O657.15[理学—分析化学] Q503[理学—化学]