Disposable electrochemical genosensor for the simultaneous analysis of different bacterial food contaminants

被引:99
作者
Farabullini, F. [1 ]
Lucarelli, F. [1 ]
Palchetti, I. [1 ]
Marrazza, G. [1 ]
Mascini, M. [1 ]
机构
[1] Univ Florence, Dipartimento Chim, I-50019 Sesto Fiorentino, FI, Italy
关键词
electrochemical genosensor; screen-printed array; DNA hybridisation; PCR; bacteria;
D O I
10.1016/j.bios.2006.06.001
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
This paper deals with the use of an electrochemical genosensor array for the rapid and simultaneous detection of different food-contaminating pathogenic bacteria. The method includes PCR amplification followed by analysis of the amplicons by hybridisation with toxin-specific oligonucleotide probes. A screen-printed array of four gold electrodes, modified using thiol-tethered oligonucleotide probes, was used. Unmodified PCR products were captured at the sensor interface via sandwich hybridisation with surface-tethered probes and biotinylated signaling probes. The resulting biotinylated hybrids were coupled with a streptavidin-alkaline phosphatase conjugate and then exposed to an a-naphthyl phosphate solution. Differential pulse voltammetry was finally used to detect the a-naphthol oxidation signal. Mixtures of DNA samples from different bacteria were detected at the nanomolar level without any cross-interference. The selectivity of the assay was also confirmed by the analysis of PCR products unrelated to the immobilised probes. (c) 2006 Elsevier B.V. All rights reserved.
引用
收藏
页码:1544 / 1549
页数:6
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