DNA microdevice for electrochemical detection of Escherichia coli O157:H7 molecular markers

被引:33
作者
Berganza, J.
Olabarria, G.
Garcia, R.
Verdoy, D.
Rebollo, A.
Arana, S.
机构
[1] GAIKER Technoll Ctr IK4 Res Alliance, Zamudio 48170, Spain
[2] CEIT, San Sebastian 20018, Spain
[3] Tecnun, San Sebastian 20018, Spain
关键词
biosensors; DNA; self-assembled monolayers; hybridization; E. coli O157 : H7;
D O I
10.1016/j.bios.2006.09.028
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
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) O157: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. (c) 2006 Elsevier B.V. All rights reserved.
引用
收藏
页码:2132 / 2137
页数:6
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