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Genetic techniques to detect E. coli 0157 - FS1211

Extremely sensitive techniques will be developed for the detection of E. coli 0157, and other verocytotoxic E. coli i foods of animal origin (meat, meat products, milk and dairy products) using the polymerase chain reaction, PCR and related techniques. Two approaches will be used, one to detect specifically E.coli 0157 and the other to detect the genes for verocytotoxin production. Having developed suitable methods, assays will be performed on a) a collection of E. coli strains to determine specificity and sensitivity, b) spiked food samples and c) actual food samples.
1. To define and sequence DNA fragments which are unique to E.coli 0157. 2. To develop a PCR specific for 0157. 3. To apply the 0157 specific detection systems using methods and samples outlined in 4 and 5 below. 4. To apply non-radiolabelled probing systems such as the combined PCR-ELISA to confirm specificity of amplification product. 5. To enhance the sensitivity and specificity of existing PCR techniques for the detection of VTEC. 6. To establish methods for the handling of food samples for the optimal release of target DNA. 7. To evaluate the enhanced PCR for the detection of 0157 and other VTEC in food samples in both spiked and actual purchased foods. 8. To consider at stage of the project the opportunity to protect intellectual property rights by patent.
Time-Scale and Cost
From: 1993

To: 1996

Cost: £66,345
Contractor / Funded Organisations
Veterinary Laboratories Agency