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用于电化学检测大肠杆菌O157:H7分子标记物的DNA微器件

DNA microdevice for electrochemical detection of Escherichia coli 0157:H7 molecular markers.

作者信息

Berganza J, Olabarria G, García R, Verdoy D, Rebollo A, Arana S

机构信息

GAIKER Technological Center-IK4 Research Alliance, Parque Tecnológico, Ed. 202. (48.170) Zamudio, Spain.

出版信息

Biosens Bioelectron. 2007 Apr 15;22(9-10):2132-7. doi: 10.1016/j.bios.2006.09.028. Epub 2006 Nov 2.

DOI:10.1016/j.bios.2006.09.028
PMID:17084071
Abstract

An electrochemical DNA sensor based on the hybridization recognition of a single-stranded DNA (ssDNA) probe immobilized onto a gold electrode to its complementary ssDNA is presented. The DNA probe is bound on gold surface electrode by using self-assembled monolayer (SAM) technology. An optimized mixed SAM with a blocking molecule preventing the nonspecific adsorption on the electrode surface has been prepared. In this paper, a DNA biosensor is designed by means of the immobilization of a single stranded DNA probe on an electrochemical transducer surface to recognize specifically Escherichia coli (E. coli) 0157:H7 complementary target DNA sequence via cyclic voltammetry experiments. The 21 mer DNA probe including a C6 alkanethiol group at the 5' phosphate end has been synthesized to form the SAM onto the gold surface through the gold sulfur bond. The goal of this paper has been to design, characterise and optimise an electrochemical DNA sensor. In order to investigate the oligonucleotide probe immobilization and the hybridization detection, experiments with different concentration of DNA and mismatch sequences have been performed. This microdevice has demonstrated the suitability of oligonucleotide Self-assembled monolayers (SAMs) on gold as immobilization method. The DNA probes deposited on gold surface have been functional and able to detect changes in bases sequence in a 21-mer oligonucleotide.

摘要

本文介绍了一种基于固定在金电极上的单链DNA(ssDNA)探针与其互补ssDNA杂交识别的电化学DNA传感器。DNA探针通过自组装单分子层(SAM)技术结合在金表面电极上。制备了一种优化的混合SAM,其中含有一种阻断分子,可防止电极表面的非特异性吸附。本文通过将单链DNA探针固定在电化学传感器表面,利用循环伏安法实验特异性识别大肠杆菌(E. coli)0157:H7互补靶DNA序列,设计了一种DNA生物传感器。已合成了在5'磷酸端含有C6烷硫醇基团的21聚体DNA探针,通过金硫键在金表面形成SAM。本文的目标是设计、表征和优化一种电化学DNA传感器。为了研究寡核苷酸探针的固定和杂交检测,进行了不同浓度DNA和错配序列的实验。该微型器件已证明寡核苷酸自组装单分子层(SAMs)在金上作为固定方法的适用性。沉积在金表面的DNA探针具有功能性,能够检测21聚体寡核苷酸中碱基序列的变化。

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