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Investigation of the compatability of complex food matrices with pathogens detection using PCR. - FS1212

Description
The project aims to produce a multiplex PCR based screening assay for the simultaneous detection of pathogens of particluar concern to MAFF in a variety of foodstuffs. We intend to concentrate primarily on overcoming the problems associated with direct PCR from complex food matrices and thereby establish and optimise a method for pathogen target DNA amplification from food samples, which minimises the sample preparation and assay time and would be broadly applicable to a defined set of foodstuffs. We aim to develop a multiplex PCR employing primers specific to the pathogen sequences of interest differentially labelled with fluorophors enabling simultaneous total microbial load assessment and detection of specific target pathogens in a single sample. To enable routine analysis, we aim to develop the assay into a simple, conventionally acceptable, microtitre format with a fluorimetric based detection system for the amplified target pathogen DNA.
Objective
1. To overcome/eliminate the problems associated with direct PCR from complex food matrices and thereby establish and optimise a method for pathogen target DNA amplification from food samples. 2. To produce a multiplex PCR based screening assay for the simultaneous detection of total microbial load, Listeria spp. , Salmonella spp. and enterotoxigenic E. coli in a variety of foodstuffs. 3. To develop the assay into a simple, conventionally acceptable, user friendly microtitre format with a fluorimetric based detection system for the amplified target pathogen DNA, in order to ensure that the assay would be suitable for routine analysis.
Time-Scale and Cost
From: 1993

To: 1996

Cost: £220,000
Contractor / Funded Organisations
LGC
Keywords