An Internalin A Probe-Based Genosensor for Listeria monocytogenes Detection and Differentiation

被引:3
|
作者
Bifulco, Laura [1 ]
Ingianni, Angela [1 ]
Pompei, Raffaello [1 ]
机构
[1] Univ Cagliari, Dept Biomed Sci, Microbiol Sect, I-09124 Cagliari, Italy
关键词
ELECTROCHEMICAL DNA BIOSENSOR; SELF-ASSEMBLED MONOLAYERS; TO-EAT FOODS; METHYLENE-BLUE; GOLD ELECTRODES; PREGNANCY; SURVEILLANCE; INVASION; ENGLAND; PROTEIN;
D O I
10.1155/2013/640163
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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.
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页数:6
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