共 15 条
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.
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页码:1544 / 1549
页数:6
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