Sensitive and cost-effective molecular methods, such as PCR, are receiving increasing attention for testing the microbiological safety of food. However, lack of international validation, standard protocols, reagents and equipment has hampered its implementation in routine diagnostic. The overall objective of the proposed project is to facilitate implementation of diagnostic PCR for detection of food-borne pathogens. This will be achieved through construction of DNA sample library and primer databank, validation of thermocyclers, ring-trials, automation and guidelines. The project will involve 12 expert and 22 end-user laboratories from 20 EU and applicant countries. The work is planned in 2 phases, including 6 WP and 20 tasks, which will focus on 5 major pathogens (Salmonella, Campylobacter, enterohemorrhagic E. coli (EHEC), L. monocytogenes and Y. enterocolitica) and 4 sample types (poultry carcass-rinse, pig carcass swab, meat and milk).
Recognising that sensitive and more cost-effective methods are needed for detecting food-borne pathogens, we are launching a project seeking to validate and standardise use of the polymerase chain reaction (PCR) for this purpose. Although a powerful research tool, the application of PCR for detecting food-borne pathogens is hampered from lack of validation, standard protocols, reagents, and equipment. Additional specific project objectives include validating a simple method for purifying DNA from bacterial cultures, establishing a central collection of certified DNA sample materials, establishing a databank containing key food-pathogen DNA sequence, listing strains for specificity testing, developing standardised reagents, and validating pre-PCR sample treatment methods
- Validation of a simple and reproducible method for purification of DNA from bacterial cultures
- DNA-sample bank, consisting of defined DNA material
- DNA databank for registration of food pathogen-specific genes primers and probes
- Sets of criteria for validation of thermocyclers
- User-friendly, pre-PCR sample preparation techniques for the sample types included in this study
- Assessment of specificity, sensitivity and reproducibility of known PCR analyses through comparative studies and ring-trials
- Validated automated, closed-tube PCR for detection of Campylobacter spp.
- Guidelines for first-time implementation of PCR by end-users
- Proposals for European standardisation (CEN/TC 275) of PCR testing
- Organisation of two workshops for technology transfer