An electrochemical competitive biosensor for ochratoxin A based on a DNA biotinylated aptamer

被引:155
作者
Bonel, Laura [1 ]
Vidal, Juan C. [1 ]
Duato, Patricia [1 ]
Castillo, Juan R. [1 ]
机构
[1] Univ Zaragoza, Inst Environm Sci IUCA, Analyt Spect & Sensors Grp, E-50009 Zaragoza, Spain
关键词
Ochratoxin A; Aptamer; Electrochemical biosensor; Screen printed; Mycotoxins; POLYCLONAL ANTIBODIES; SMALL MOLECULES; ASSAY; IMMUNOSENSORS; APTASENSOR; FOOD;
D O I
10.1016/j.bios.2010.12.036
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
Ochratoxin A (OTA) is one of the most important mycotoxin contaminants of foods, particularly cereals and cereal products, with strict low regulatory levels (of ppb) in many countries worldwide. An electrochemical competitive aptamer-based biosensor for OTA is described. Paramagnetic microparticle beads (MBs) were functionalized with an aptamer specific to OTA, and were allowed to compete with a solution of the mycotoxin conjugated to the enzyme horseradish peroxidase (OTA-HRP) and free OTA. After separation and washing steps helped with magnetic separations, the modified MBs were localized on disposable screen-printed carbon electrodes (SPCEs) under a magnetic field, and the product of the enzymatic reaction with the substrate was detected with differential-pulse voltammetry. In addition to magnetic separation assays, other competitive schemes (direct/indirect aptasensors performed on the SPCEs surface or using gold nanoparticles functionalized with the aptamer) were preliminary tested, optimized and compared. The magnetic aptasensor showed a linear response to OTA in the range 0.78-8.74 ng mL(-1) and a limit of detection of 0.07 +/- 0.01 ng mL(-1), and was accurately applied to extracts of certified and spiked wheat samples with an RSD lower than about 8%. (C) 2010 Elsevier B.V. All rights reserved.
引用
收藏
页码:3254 / 3259
页数:6
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