NDLI: Detection of hlyA Gene of Listeria Monocytogenes with Electrochemical DNA Biosensor

Content Provider	IEEE Xplore Digital Library
Author	Lingwei Wu Quanjun Liu Zhongwei Wu Zuhong Lu
Copyright Year	2008
Description	Author affiliation: State key Lab. of Bioelectronics, Southeast Univ., Nanjing (Lingwei Wu)
Abstract	Listeria monocytogenes (LM) is a bacterial foodborne pathogen responsible for listeriosis, an illness characterized by encephalitis, septicaemia , and meningitis. One of the best ways to detect and confrim the pathogen is through the detection of one of the virulence factors, listeriolysin O (LLO) produced by the microorganism. LLO is absolutely required for virulence by L. monocytogenes, and is found only in virulent strains of the species. This paper focuses on the electrical detection methods used to detect the LLO toxin gene in food products or organism. A new and simple electrochemical approach for hybridisation detection without the labelling the target DNA is described. The coupling of DNA electrochemical sensors have specialities of quick detection and cheapness and have the potential of the quantitative evaluation of the hlyA gene of Listeria monocytogenes. The electrochemical sensor was obtained by immobilizing single-stranded oligonucleotides onto gold electrodes through mercaptan of the DNA bases using N- hydroxysulfosuccinimide (NHS) and N-(3-dimethylamion)propyl-N'-ethyl carbodiimidehydrochloride (EDC) as activation. The hybridization reaction that occurred on the electrode surface was evidenced by cyclic voltammetry (CV) analysis - using as $[Co(phen)_{3}](C1O_{4})_{3}$ indicator. The covalently immobilized single-stranded DNA could selectively hybridize with its complementary DNA in solution to form double-stranded DNA on the gold surface. A significant increase of the peak current of cyclic voltammetry (CV) upon the hybridization of immobilized ssDNA with PCR amplification products in the solution was observed. This peak current change was used to monitor the amount of PCR amplification products. Factors such as DNA target concentration and hybridization conditions determining the sensitivity of the electrochemical assay were investigated.
Starting Page	375
Ending Page	378
File Size	118009
Page Count	4
File Format	PDF
ISBN	9781424417476
DOI	10.1109/ICBBE.2008.95
Language	English
Publisher	Institute of Electrical and Electronics Engineers, Inc. (IEEE)
Publisher Date	2008-05-16
Publisher Place	China
Access Restriction	Subscribed
Rights Holder	Institute of Electrical and Electronics Engineers, Inc. (IEEE)
Subject Keyword	Electrodes Pathogens Gold Microorganisms DNA Capacitive sensors Organisms Food products Biosensors Labeling
Content Type	Text
Resource Type	Article

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