Zhu Ningning, Lin Yuqing, Yu Ping, Su Lei, Mao Lanqun
Beijing National Laboratory for Molecular Sciences, Institute of Chemistry, The Chinese Academy of Sciences, Beijing 100190, China.
Anal Chim Acta. 2009 Sep 14;650(1):44-8. doi: 10.1016/j.aca.2009.05.017. Epub 2009 May 21.
This study describes a simple and label-free electrochemical impedance spectroscopic (EIS) method for sequence-specific detection of DNA by using single-walled carbon nanotubes (SWNTs) as the support for probe DNA. SWNTs are confined onto gold electrodes with mixed self-assembly monolayers of thioethanol and cysteamine. Single-stranded DNA (ssDNA) probe is anchored onto the SWNT support through covalent binding between carboxyl groups at the nanotubes and amino groups at 5' ends of ssDNA. Hybridization of target DNA with the anchored probe DNA greatly increases the interfacial electron-transfer resistance (R(et)) at the double-stranded DNA (dsDNA)-modified electrodes for the redox couple of Fe(CN)(6)(3-/4-), which could be used for label-free and sequence-specific DNA detection. EIS results demonstrate that the utilization of SWNTs as the support for probe DNA substantially increases the surface loading of probe DNA onto electrode surface and thus remarkably lowers the detection limit for target DNA. Under the conditions employed here, R(et) is linear with the concentration of target DNA within a concentration range from 1 to 10 pM with a detection limit down to 0.8 pM (S/N=3). This study may offer a novel and label-free electrochemical approach to sensitive sequence-specific DNA detection.
本研究描述了一种简单且无需标记的电化学阻抗谱(EIS)方法,用于通过使用单壁碳纳米管(SWNTs)作为探针DNA的载体来进行DNA的序列特异性检测。SWNTs通过硫代乙醇和半胱胺的混合自组装单分子层被限制在金电极上。单链DNA(ssDNA)探针通过纳米管上的羧基与ssDNA 5'端的氨基之间的共价结合锚定在SWNT载体上。目标DNA与锚定的探针DNA杂交极大地增加了双链DNA(dsDNA)修饰电极上对于Fe(CN)(6)(3-/4-)氧化还原对的界面电子转移电阻(R(et)),这可用于无需标记的序列特异性DNA检测。EIS结果表明,利用SWNTs作为探针DNA的载体显著增加了探针DNA在电极表面的表面负载量,从而显著降低了目标DNA的检测限。在此处采用的条件下,R(et)与目标DNA浓度在1至10 pM的浓度范围内呈线性关系,检测限低至0.8 pM(S/N = 3)。本研究可能为灵敏的序列特异性DNA检测提供一种新颖的无需标记的电化学方法。