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Validation and standardization of diagnostic Polymerase Chain Reaction
for detection of food-borne pathogens
FOOD-PCR

Contract number : QLK1-1999-00226
Contract type : Shared Cost Project
Total cost : € 2.326.333
EC contribution : € 1.607.220
Starting date : 1/04/2000
Duration : 36 Months
Scientific Officer : Alkmini Katsada
Project website : not yet available
Coordinator
Dr Jeffrey Hoorfar
Danish Veterinary Laboratory
Department of Microbiology
27, Bülowsvej
1790 Copenhagen
Denmark
Tel.: +45-35300251
Fax: +45-35300120
E-mail: www.pcr.dk
Background

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).

Objectives

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

(expected) Results and achievements
  • 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

Partners

Lund University
Department of Applied Microbiology
Getingevägen 60
PO Box 124
221 00 Lund
Sweden
University of Veterinary Science
Institut of Milk Hygiene, Milk Technology and Food Science
Veterinärplatz 1
1210 Wien
Austria
Institute for Reference Materials and Measurements
Retieseweg
2440 Geel
Belgium
Federal Institute for Health Protection of Consumers and Veterinary Medecine
Division Diagnosis and Epidemiology.
Diedersdorfer Weg 1
12277 Berlin
Germany
Congen Biotechnologie GmbH
Robert-Rössel Straße 10
13125 Berlin
Germany
University of Athens
Microbiology
M. Asias 75
11527 Athens
Greece
Institute of Animal Physiology
Slovak Academy of Sciences
Soltesovej 4
040 01 Kosice
Slovakia
Dr Bülte, Michael
Justus-Liebig-Universitat Gießen
Institute of Veterinarian Food Science
Frankfurter Straße 92
35392 Giessen
Germany
Agence Française de Sécurité Sanitaire des Aliments
43, Rue de Dantzig
75015 Paris
France
Universidad Complutense de Madrid
Dept. Patología Animal
Unidad de Microbiología
28040 Madrid
Spain
Danish Meat Research Institute
Process Technology and Hygiene
Maglegårdsvej 2
4000 Roskilde
Denmark
National Institute of Public Health
Centre for Food Chain Hygiene
Palackeho . 1-3
612 42 Brno
Czech Republic
Vyskumny Ustav Potravinarsky
Food Research Institute
Priemyselna 4
82475 Bratislava
Slovakia
Institute of Chemical Technology Prague
Department of Biochemistry and Microbiology
Technickä 1903/3
166 28 Prague
Czech Republic
Meat Technology Center
Granja Camps I Armet, S/N
17121 Monells
Spain
National Institute for Engineering and Industrial Technology
Departamento de Tecnología das Indústrias Alimentares
Estrada do Paco do Lumiar
Edificio S
1649-038 Lisboa
Portugal
Coop Italia
Via Del Lavoro 6/8
40033 Casalecchio di Bologna
Italy
Instituto Superiore di Sanità
Reparto Igiene delle Tecnologie Alimentari
Lab Alimenti
V. Le Regina Elena N. 299
00166 Roma
Italy
Medicanalysis Ltd.
Fleming 18, Maroysi
15126 Athens
Greece
Nestlé Research Center
Department Quality & Safety
PO Box 44
1000 Lausanne
Switzerland
Estonian Agrobiocentre
Laboratory of Mycobacterioses and Tuberculin
Roomu Tee 10
51013 Tartu
Estonia
Norwegian Food Research Institute Matforsk
Osloveien 1
1430 Ås
Norway
Swedish Meats r & d Ab
V. Langgatan 20
PO Box P.0.Box 504
244 24 Kävlingen
Sweden
National Food Agency
Biology Division
Hamnesplanaden 5
PO Box 622
751 26 Uppsala
Sweden


Fifth Framework Programme

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:

Volume 1 (PDF 2.9 MB)

   

Volume 2 (PDF 1.9 MB)

 

Last update

:

23-09-2003



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