Yang Jie, Jiao Kui, Yang Tao
College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042, China.
Anal Bioanal Chem. 2007 Oct;389(3):913-21. doi: 10.1007/s00216-007-1450-5. Epub 2007 Sep 13.
Carboxyl group-functionalized single-walled carbon nanotubes (SWNTs) and 2,6-pyridinedicarboxylic acid (PDC) were electropolymerized by cyclic voltammetry on a glassy-carbon electrode (GCE) surface to form composite films (SWNTs/PDC). Zirconia was then electrodeposited on the SWNTs/PDC/GCE from an aqueous electrolyte containing ZrOCl2 and KCl by cycling the potential between -1.1 V and +0.7 V at a scan rate of 20 mV s(-1). DNA probes with a phosphate group at the 5' end were easily immobilized on the zirconia thin films, because of the strong affinity between zirconia and phosphate groups. The sensors were characterized by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). EIS was used for label-free detection of the target DNA by measuring the increase of the electron transfer resistance (R(et)) of the electrode surface after the hybridization of the probe DNA with the target DNA. The PAT gene fragment and polymerase chain reaction (PCR) amplification of the NOS gene from transgenically modified beans were satisfactorily detected by use of this DNA electrochemical sensor. The dynamic range of detection of the sensor for the PAT gene fragment was from 1.0 x 10(-11) to 1.0 x 10(-6) mol L(-1) and the detection limit was 1.38 x 10(-12) mol L(-1).
通过循环伏安法在玻碳电极(GCE)表面将羧基功能化的单壁碳纳米管(SWNTs)和2,6 - 吡啶二甲酸(PDC)进行电聚合,以形成复合膜(SWNTs/PDC)。然后通过在含有ZrOCl2和KCl的水性电解质中,以20 mV s(-1)的扫描速率在-1.1 V和+0.7 V之间循环电势,将氧化锆电沉积在SWNTs/PDC/GCE上。由于氧化锆与磷酸基团之间具有很强的亲和力,5'端带有磷酸基团的DNA探针很容易固定在氧化锆薄膜上。通过循环伏安法(CV)和电化学阻抗谱(EIS)对传感器进行了表征。EIS用于通过测量探针DNA与目标DNA杂交后电极表面电子转移电阻(R(et))的增加来对目标DNA进行无标记检测。使用这种DNA电化学传感器对转基因豆中的PAT基因片段和NOS基因的聚合酶链反应(PCR)扩增进行了令人满意的检测。该传感器对PAT基因片段的检测动态范围为1.0×10(-11)至1.0×10(-6) mol L(-1),检测限为1.38×10(-12) mol L(-1)。