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Biomed Res Int. 2013;2013:640163. doi: 10.1155/2013/640163. Epub 2013 Mar 20.

An internalin a probe-based genosensor for Listeria monocytogenes detection and differentiation.

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1
Department of Biomedical Sciences, Section of Microbiology, University of Cagliari, via Porcell 4, 09124 Cagliari, Italy.

Abstract

Internalin A (InlA), a protein required for Listeria monocytogenes virulence, is encoded by the inlA gene, which is only found in pathogenic strains of this genus. One of the best ways to detect and confirm the pathogenicity of the strain is the detection of one of the virulence factors produced by the microorganism. This paper focuses on the design of an electrochemical genosensor used to detect the inlA gene in Listeria strains without labelling the target DNA. The electrochemical sensor was obtained by immobilising an inlA gene probe (single-stranded oligonucleotide) on the surfaces of screen-printed gold electrodes (Au-SPEs) by means of a mercaptan-activated self-assembled monolayer (SAM). The hybridisation reaction occurring on the electrode surface was electrochemically transduced by differential pulse voltammetry (DPV) using methylene blue (MB) as an indicator. The covalently immobilised single-stranded DNA was able to selectively hybridise to its complementary DNA sequences in solution to form double-stranded DNA on the gold surface. A significant decrease of the peak current of the voltammogram (DPV) upon hybridisation of immobilised ssDNA was recorded. Whole DNA samples of L. monocytogenes strains could be discriminated from other nonpathogenic Listeria species DNA with the inlA gene DNA probe genosensor.

PMID:
23586053
PMCID:
PMC3618917
DOI:
10.1155/2013/640163
[Indexed for MEDLINE]
Free PMC Article
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