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) (KAR id:71942)
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. | |
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: | Divisions > Division of Natural Sciences > Physics and Astronomy |
Funders: |
EC Systems (Poland) (https://ror.org/03d73se48)
Organisations -1 not found. Organisations -1 not found. |
Depositing User: | Ian Bruce |
Date Deposited: | 01 Feb 2019 16:35 UTC |
Last Modified: | 05 Nov 2024 12:34 UTC |
Resource URI: | https://kar.kent.ac.uk/id/eprint/71942 (The current URI for this page, for reference purposes) |
- Export to:
- RefWorks
- EPrints3 XML
- BibTeX
- CSV
- Depositors only (login required):