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A multiplex magnetic capture hybridisation and multiplex Real-Time PCR protocol for pathogen detection in seafood

Amagliani, G., Omiccioli, E., Brandi, G., Bruce, Ian J., Magnani, M. (2010) A multiplex magnetic capture hybridisation and multiplex Real-Time PCR protocol for pathogen detection in seafood. Food Microbiology, 27 (5). pp. 580-585. ISSN 0740-0020. (doi:10.1016/j.fm.2010.01.007) (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided)

The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided. (Contact us about this Publication)
Official URL
http://dx.doi.org/10.1016/j.fm.2010.01.007

Abstract

Seafood could become a source of bacterial pathogens by exposure to contaminated water or through processing practices, thus representing a public health hazard. Conventional culture-based analytical methods take several days to be completed, while the molecular rapid identification of bacterial pathogens is crucial for effective disease control. The developed application consist of a multiplex magnetic capture hybridisation (mMCH) assay for the simultaneous isolation of Salmonella spp. and Listeria monocytogenes DNA from seafood, using paramagnetic amino-modified nanoparticles with capture oligonucleotides, and a triplex Real-Time PCR with an Internal Amplification Control (IAC), in accordance with ISO 22174. The detection probability was 100 with 10 genome equivalents of each target species co-amplified in the same reaction. The complete molecular procedure was tested on raw and smoked salmon fillets artificially contaminated with known amounts of one or both target bacteria (1-103cfu/g), directly or after culture enrichment, and compared for equivalence with the standard methods. Results revealed a complete agreement between the two approaches, with a sensitivity of 1cfu/g, in enriched samples, and higher sensitivity (102-103cfu/g) of the molecular method in samples examined before culture enrichment. The proposed procedure was also able to identify a natural contamination by L. monocytogenes in smoked salmon with a considerable shortening of time.

Item Type: Article
DOI/Identification number: 10.1016/j.fm.2010.01.007
Uncontrolled keywords: animal; article; evaluation; food contamination; genetics; isolation and purification; Listeria monocytogenes; magnetism; methodology; microbiology; nucleic acid hybridization; polymerase chain reaction; salmon; Salmonella; sea food, Animals; Food Contamination; Listeria monocytogenes; Magnetics; Nucleic Acid Hybridization; Polymerase Chain Reaction; Salmon; Salmonella; Seafood, Bacteria (microorganisms); Listeria monocytogenes; Salmonella
Subjects: Q Science
Q Science > QH Natural history
Q Science > QH Natural history > QH301 Biology
Divisions: Faculties > Sciences > School of Physical Sciences
Faculties > Sciences > School of Physical Sciences > Functional Materials Group
Depositing User: I. Bruce
Date Deposited: 01 Feb 2019 16:35 UTC
Last Modified: 30 May 2019 08:49 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/71942 (The current URI for this page, for reference purposes)
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