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基于纳米金修饰离子液体修饰碳糊电极的自组装膜的电化学脱氧核糖核酸生物传感器。

Electrochemical deoxyribonucleic acid biosensor based on the self-assembly film with nanogold decorated on ionic liquid modified carbon paste electrode.

机构信息

Shandong Entry-Exit Inspection and Quarantine Bureau of the People's Republic of China, Qingdao, China.

出版信息

Anal Chim Acta. 2011 Oct 17;704(1-2):133-8. doi: 10.1016/j.aca.2011.07.044. Epub 2011 Aug 9.

Abstract

An electrochemical DNA biosensor was fabricated by self-assembling probe single-stranded DNA (ssDNA) with a nanogold decorated on ionic liquid modified carbon paste electrode (IL-CPE). IL-CPE was fabricated using 1-butylpyridinium hexafluorophosphate as the binder and the gold nanoparticles were electrodeposited on the surface of IL-CPE (Au/IL-CPE). Then mercaptoacetic acid was self-assembled on the Au/IL-CPE to obtain a layer of modified film, and the ssDNA probe was further covalently-linked with mercaptoacetic acid by the formation of carboxylate ester with the help of N-(3-dimethylamino-propyl)-N'-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide. The hybridization reaction with the target ssDNA was monitored with methylene blue (MB) as the electrochemical indicator. Under the optimal conditions, differential pulse voltammetric responses of MB was proportional to the specific ssDNA arachis sequences in the concentration range from 1.0×10(-11) to 1.0×10(-6) mol L(-1) with the detection limit as 1.5×10(-12) mol L(-1) (3σ). This electrochemical DNA sensor exhibited good stability and selectivity with the discrimination ability of the one-base and three-base mismatched ssDNA sequences. The polymerase chain reaction product of arachis Arabinose operon D gene was successfully detected by the proposed method, which indicated that the electrochemical DNA sensor designed in this paper could be further used for the detection of specific ssDNA sequence.

摘要

一种电化学 DNA 生物传感器是通过将纳米金修饰在离子液体修饰的碳糊电极(IL-CPE)上的探针单链 DNA(ssDNA)自组装而成的。IL-CPE 是使用 1-丁基吡啶六氟磷酸盐作为粘合剂制备的,并且金纳米粒子被电沉积在 IL-CPE 的表面上(Au/IL-CPE)。然后,巯基乙酸自组装在 Au/IL-CPE 上,以获得修饰层,并且 ssDNA 探针通过 N-(3-二甲氨基丙基)-N'-乙基碳二亚胺盐酸盐和 N-羟基琥珀酰亚胺的形成与巯基乙酸进一步共价连接,形成羧酸酯。通过将亚甲基蓝(MB)作为电化学指示剂来监测与靶 ssDNA 的杂交反应。在最佳条件下,MB 的差分脉冲伏安响应与特定 ssDNA 花生序列在 1.0×10(-11)至 1.0×10(-6)mol L(-1)浓度范围内呈正比,检测限为 1.5×10(-12)mol L(-1)(3σ)。该电化学 DNA 传感器具有良好的稳定性和选择性,并且具有区分单碱基和三碱基错配 ssDNA 序列的能力。通过所提出的方法成功检测到花生阿拉伯糖操纵子 D 基因的聚合酶链反应产物,这表明本文设计的电化学 DNA 传感器可以进一步用于检测特定的 ssDNA 序列。

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