Electrochemical detection of foodborne pathogen Aeromonas hydrophila by DNA hybridization biosensor
Electrochemical detection of foodborne pathogen Aeromonas hydrophila by DNA hybridization biosensor
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DOI：
10.1016/j.bios.2010.08.030
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发表时间：
2010-12-15
影响因子：
12.6
通讯作者：
Filipiak, M.
中科院分区：
文献类型：
--
作者：
Tichoniuk, M.;Gwiazdowska, D.;Filipiak, M.
报道了一种用于检测嗜水气单胞菌的电化学DNA生物传感器。这种机会致病菌被认为是一种新兴的食源性危害，并与多种毒力因子相关，包括产生细胞毒性肠毒素气溶素。基因传感器识别层是使用由硫醇化单链DNA探针（ssDNA）和稀释剂分子-巯基醇：巯基乙醇（MCE）或巯基己醇（MCH）或巯基壬醇（MCN）组成的混合自组装单层（SAM）制备的。伏安法检测的生物传感器识别界面的双层电容支持计时库仑定量的DNA存在于电极表面上表明，混合的ssDNA和MCH单层显示最低的缺陷。其双电层电容为4.0 μ F·cm ~（-2），ssDNA探针的表面覆盖率达到8.5 × 10 ~（11）分子·cm ~（-2）。采用计时库仑法定量DNA和方波伏安法（SWV）测量电活性指示剂亚甲基蓝（MB），研究了杂交反应时间、靶DNA片段浓度和非互补DNA的存在对基因传感器识别界面电化学响应的影响.该生物传感器能够区分从A.大肠杆菌（浓度为2.5 μ g cm（-3））和其他微生物DNA。(C)2010 Elsevier B. V.保留所有权利。
The paper describes an electrochemical DNA biosensor used for the detection of Aeromonas hydrophila. This opportunistic pathogen is recognized as an emerging foodborne hazard and is associated with a variety of virulence factors including production of cytotoxic enterotoxin aerolysin. The genosensor recognition layer was prepared using mixed self-assembled monolayer (SAM) consisting of thiolated single-stranded DNA probe (ssDNA) and diluent molecules - mercaptoalcohol: mercaptoethanol (MCE) or mercaptohexanol (MCH) or mercaptononanol (MCN). The voltammetric examination of double-layer capacitance of biosensor recognition interface supported by chronocoulometric quantitation of DNA present on the electrode surface showed that mixed ssDNA and MCH monolayer revealed the lowest defectiveness. Its double-layer capacitance equaled 4.0 mu F cm(-2) and ssDNA probe surface coverage reached 8.5 x 10(11) molecules cm(-2) of gold electrode surface. Chronocoulometric quantitation of DNA and square wave voltammetry (SWV) measurements of electroactive indicator, methylene blue (MB) were performed to investigate the influence of hybridization reaction time, concentration of target DNA fragments, and presence of non-complementary DNA on the electrochemical response of genosensor recognition interface. The biosensor enabled distinction between the DNA samples isolated from A. hydrophila (present at the concentration of 2.5 mu g cm(-3)) and other microbial DNA. (C) 2010 Elsevier B.V. All rights reserved.