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基于主体-客体识别技术的均相杂交非固定化电化学生物传感策略。

A non-immobilizing electrochemical DNA sensing strategy with homogenous hybridization based on the host-guest recognition technique.

机构信息

Department of Chemistry, East China Normal University, 3663, Zhongshan Road (N), Shanghai, China.

出版信息

Biosens Bioelectron. 2011 Jan 15;26(5):2655-9. doi: 10.1016/j.bios.2010.02.021. Epub 2010 Mar 1.

DOI:10.1016/j.bios.2010.02.021
PMID:20346643
Abstract

In this paper, we report a new strategy for electrochemical DNA detection in homogeneous solution based on the host-guest molecule recognition technique. In this sensing protocol, a novel dually labeled DNA probe (DLP) in a stem-loop structure was employed, which was designed with dabcyl labeled at one end as a guest molecule, and with Au nanoparticle labeled at the other end as electrochemical tag to indicate the hybridization occurrence. One α-CD/MCNTs/GCE was used for capturing the DNA hybridization and electrochemical signal transduction. Before the hybridization, the DLP remained in the stem-loop structure, which forced the dabcyl molecular to be closed to the Au nanoparticle. Due to the steric effect of the Au nanoparticle, the dabcyl was prevented from conjugating with the α-CD on the electrode and resulting in that the DLP could not be captured by the electrode. After hybridized with the target DNA, the target-binding DLP caused the DLP's loop-stem structure opened and then the dabcyl molecule was easily entering the cavity of the α-CD modified electrode and resulting in that the DLP could be captured by the α-CD modified electrode and the capture efficiency was proportion with the concentration of the target DNA. Therefore, the target hybridization event can be sensitively transduced via detecting the electrochemical reduction current signal of AuCl(4)(-) of Au nanoparticles labeled at the DLP. Using this strategy, as low as 2.6×10(-10) M DNA target had been detected with excellent differentiation ability for even single mismatch.

摘要

在本文中,我们报告了一种基于主体-客体分子识别技术的均相溶液中电化学 DNA 检测的新策略。在这种传感方案中,采用了一种新型的茎环结构的双重标记 DNA 探针(DLP),其一端标记有 dabcyl 作为客体分子,另一端标记有金纳米粒子作为电化学标记物,以指示杂交发生。一个 α-CD/MCNTs/GCE 用于捕获 DNA 杂交和电化学信号转导。在杂交之前,DLP 保持在茎环结构中,迫使 dabcyl 分子靠近金纳米粒子。由于金纳米粒子的空间位阻效应,dabcyl 被阻止与电极上的 α-CD 结合,导致 DLP 不能被电极捕获。与靶 DNA 杂交后,靶结合的 DLP 导致 DLP 的环-茎结构打开,然后 dabcyl 分子容易进入 α-CD 修饰电极的空腔,导致 DLP 可以被 α-CD 修饰电极捕获,并且捕获效率与靶 DNA 的浓度成正比。因此,通过检测 DLP 标记的金纳米粒子的 AuCl(4)(-)的电化学还原电流信号,可以灵敏地转导靶杂交事件。利用这种策略,甚至可以检测到低至 2.6×10(-10) M 的 DNA 靶标,并且具有出色的区分能力,即使是单个错配。

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