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Food-PCR: Validation and standardisation of diagnostic Polymerase Chain Reaction for detection of foodborne pathogens

Abstract
The overall objective of the proposal is to facilitate implementation of diagnostic Polymerase Chain reaction (PCR) for both verfication and detection of foodborne pathogens through harmonization and standardization of methods in Europe. It is known that PCR testing, despite enormous diagnostic potential, has had difficulty moving from research into end-user laboratories. Part of the problem is technical, but lack of international validation, the need for training and difficulties in transfer of technology, also play major roles.
The project deals with five major zoonotic pathogens: Salmonella spp. , Campylobacter spp., EHEC, Listeria monocytogenes, Yersinia enterocolitica.
The objectives are:
1. to validate a simple method for purification of DNA from bacterial culture
2. to establish a DNA bank
3. to establish a databank
4. to develop measureable criteria for validation of thermocyclers;
5. to develop pre-PCR sample preparation techniques
6. to assess specifity, sensitivity and reproducibility of known PCR analyses
7. to develop practical guidelines for the first-time implementation of PCR by end-users
8. to prepare draft proposals for European standardization (CEN)
9. to arrange hands-on workshops for transfer of technology
Coordination for vetmeduni vienna
Schauberger Günther
Duration
01.03.00-28.02.03
Type of Research
Applied research

Further information: http://www.pcr.dk

Staff
Schauberger G.,
Vetmed Research Units
Institute of Physiology, Pathohysiology and Biophysics, Unit of Physiology and Biophysics
Funded by
EU (Kommission der Europäischen Union), Brussels, European Union
15 Publications

Schoder, D; Schmalwieser, A; Schauberger, G; Hoorfar, J; Kuhn, M; Wagner, M (2005): Novel approach for assessing performance of PCR cyclers used for diagnostic testing. J Clin Microbiol. 2005; 43(6):2724-2728
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Josefsen, MH; Cook, N; D"Agostino, M; Hansen, F; Wagner, M; Demnerova, K; Heuvelink, AE; Tassios, PT; Lindmark, H; Kmet, V; Barbanera, M; Fach, P; Loncarevic, S; Hoorfar, J (2004): Validation of a PCR-based method for detection of food-borne thermotolerant campylobacters in a multicenter collaborative trial. Appl Environ Microbiol. 2004; 70(7):4379-4383
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D"Agostino, M; Wagner, M; Vazquez-Boland, JA; Kuchta, T; Karpiskova, R; Hoorfar, J; Novella, S; Scortti, M; Ellison, J; Murray, A; Fernandes, I; Kuhn, M; Pazlarova, J; Heuvelink, A; Cook, N (2004): A validated PCR-based method to detect Listeria monocytogenes using raw milk as a food model--towards an international standard. J Food Prot. 2004; 67(8):1646-1655

Hoorfar, J; Malorny, B; Abdulmawjood, A; Cook, N; Wagner, M; Fach, P (2004): Practical considerations in design of internal amplification controls for diagnostic PCR assays. J Clin Microbiol. 2004; 42(5):1863-1868
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Lübeck, PS; Cook, N; Wagner, M; Fach, P; Hoorfar, J (2003): Toward an international standard for PCR-based detection of food-borne thermotolerant Campylobacters: validation in a multicenter collaborative trial. Appl Environ Microbiol. 2003; 69(9):5670-5672
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Malorny, B; Tassios, PT; Rådström, P; Cook, N; Wagner, M; Hoorfar, J (2003): Standardization of diagnostic PCR for the detection of foodborne pathogens. Int J Food Microbiol. 2003; 83(1):39-48

Schoder, D; Schmalwieser, A; Schauberger, G; Kuhn, M; Hoorfar, J; Wagner, M (2003): Physical characteristics of six new thermocyclers. Clin Chem. 2003; 49(6 Pt 1):960-963
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Schoder, D; Schmalwieser, A; Schauberger G; Kuhn, M; Wagner, M (2002): Thermocycler Evaluierung: Überprüfung der räumlichen und zeitlichen Temperaturverteilung - Einflussnahme der physikalischen Performance auf die Amplifikationseffizienz. -28. Jahrestagung der Österreichischen Gesellschaft für Hygiene, Mikrobiologie und Präventivmedizin; MAY 7-10, 2002; Meran, Südtirol. Abstract 89;

Kuhn, M; Mergemeier, S; Schoder, D; Wagner, M (2002): Funktioniert der Thermocycler richtig? Labor-Praxis (4) 80-83.

Schauberger, G; Wagner, M (1998): Qualitätskontrolle von PCR Thermocyclern: Überprüfung der räumlichen und zeitlichen Temperaturverteilung. Vorträge der gemeinsamen Jahrestagung der Österreichischen Gesellschaft für Biomedizinische Technik und der Österreichischen Gesellschaft für Medizinische Physik, Wien, Austria, MAY 21-23, 1998. Biomedizinische Technik = Biomedical engineering (43), S2 216-218 .

Ellison, J; D´Agostino, M; Karpiskova, R; Novella, S; Sanchez, M; Schoder, D; Vasquez-Boland, J. A; Wagner, M; Cook, N (2001): Detection of Listeria monocytogenes by PCR. Second general Meeting of the Food-PCR Project, Lund University, Sweden, Sweden, June 2001. Second general Meeting of the Food-PCR Project, Lund University, Sweden, June 2001.

Kuhn, M; Mergemeier, S; Schoder, D; Wagner, M (2001): A temperaturesensitive PCR-System for the validation of thermocyclers used in diagnostic PCR. Second general Meeting of the FOOD-PCR Project, Lund University, Schweden, Sweden, June 2001. Second general Meeting of the FOOD-PCR Project, Lund University, Schweden, June 2001.

Schoder, D; Schmalwieser, A; Schauberger, G; Kuhn, M; Wagner, M (2001): Food testing by PCR: Thermal Processor Reliability Testing. Euroconference: Food safety assurance and veterinary public health; the topical research issues; Event 2: Safety assurance during food processing, Wien, Austria, April 2001. Euroconference: Food safety assurance and veterinary public health; the topical research issues; Event 2: Safety assurance during food processing, Wien, April 2001.

Schoder, D; Schmalwieser, A; Schauberger, G; Kuhn, M; Wagner, M (2002): Qualitätskontrolle von Thermocyclern: Überprüfung der physikalischen Performance und deren Einflußnahme auf die Amplifikationseffizienz. 831-835.-42. Arbeitstagung des Arbeitsgebietes Lebensmittelhygiene, Deutsche Veterinärmedizinische Gesellschaft; SEP 25-28, 2001; Garmisch-Partenkirchen.

Schoder, D; Schmalwieser, A; Schauberger, G; Kuhn, M; Wagner, M (2001): Thermal processor reliability testing. Second General Meeting of the Food-PCR Project, Lund, April 2001. Second General Meeting of the Food-PCR Project, Lund, April 2001.

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