Abstract
Food-borne zoonotic bacteria, transferred from animals to food products, are a major concern in modern food production and, consequently, for human health (1). Accordingly, there is a need to control the entire food chain, from infections at the herd level to the consumer, through screening and certification programs, which apply highly sensitive and cost-effective methods for detection of food-borne pathogens. Such tests would also be useful to the food industry and clinical diagnostic laboratories. Molecular methods, using nucleic acid diagnostics, are receiving increasing attention for testing the microbiological safety of food (2). There are several advantages of nucleic acid diagnostics compared to bacteriological culturing and immunochemical methods. Nucleic acid molecules can, via sequence combinations, precisely prescribe the phenotypic characteristics of a microorganism, and detection of specific sequences unique to a particular species can obviate any requirement for confirmatory tests. Further-more, detection of nucleic acids is very rapid compared to conventional culture-based analysis. The most widely used nucleic acid diagnostic test is based on the powerful PCR technology (3).
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References
Tauxe, R. V. (1991) Salmonella: A postmodern pathogen. J. Food Protection 54, 563–568.
Vaneechoutte, M., and Van Eldere, J. (1997) The possibilities and limitations of nucleic acid amplification technology in diagnostic microbiology. J. Med. Microbiol. 46, 188–194.
Mullis, K., Faloona, F., Scharf, S., Siaki, R., Horn, G., Erlich, H. (1986) Specific enzymatic amplification of DNA in vitro: The polymerase chain reaction. Cold Spring Harb. Symp. Quant. Biol. 51, 263–273.
Scheu P. M., Berghof K., and Stahl U. (1998) Detection of pathogenic and spoilage micro-organisms in food with the polymerase chain reaction. Food Micorbiol. 15, 13–31.
Anon. (1999 A) Microbiology of food and animal feeding stuff-carcass sampling for microbiological analysis. ISO/CD 17604. International Organization for Standardization, Geneva, Switzerland.
Anon. (2000) Microbiology of food and animal feeding stuff-Preparation of test samples, initial suspension and decimal dilution for microbiological examination. Doc CEN/TC 275/ WG 6 N 114. Published by AFNORE, Paris, France.
Anon. (1999) Microbiology of food and feeding stuffs-Protocol for the validation of alternative methods. Document established by the Joint Group of MicroVal (WG 6) and CEN (WG 6/TAG 2). ISO/DIS 16140, Geneva, Switzerland.
Anon. (2001) General requirement relating to methods of detecting foodborne pathogens using the polymerase chain reaction (PCR). CEN/TC 275/WG6/TAG3 N 10. Published by DIN. Berlin, Germany.
D’Aoust, J.-Y., Sewell, A., and Jean, A. (1990) Limited sensitivity of short (6h) selective enrichment for detection of foodborne Salmonella. J. Food Protection 53, 562–265.
Hoorfar, J. and Baggesen, D. L. (1998) Importance of preenrichment media for isolation of Salmonella spp. from swine and poultry. FEMS Microbiol. Lett. 169, 125–130.
Knutsson, R., Blixt, Y., Grage, H., Borch, E., and Rådström, P. (2002) Evaluation of selective enrichment PCR procedures for Yersinia enterocolitica. Int. J. Food Microbiol. 73, 35–46.
Uyttendaele, M., Shukkink, R., van Gemen, B., and Debevere, J. (1995) Detection of Campylobacter jejuni added to foods by using a combined selective enrichment and nucleic acid sequence-based amplification (NASBA). Appl. Environ. Microbiol. 61, 1341–1347.
Szabo, E. A., and Mackey, B. M. (1999) Detection of Salmonella enteritidis by reverse transcription-polymerase chain reaction (PCR). Int. J. Food Microbiol. 51, 113–122.
Anon. (1993) Microbiology-general guidance on methods for the detection of Salmonella. International Organization for Standardization, ISO 6579, 3rd edition. Geneva, Switzerland.
Anon. (1995) Microbiology of food and animal feeding stuffs-Horizontal method for detection of thermotolerant Campylobacter. International Standard Organisation, ISO 10272, 1st edition. Geneva, Switzerland.
Anon. (1996) Yersinia enterocolitica. Detection in food. Nordic Committee on Food Analysis, Method no. 117, 3rd Edition. Oslo, Norway.
Simon, M. C., Gray, D. I., and Cook, N. (1996) DNA extraction and PCR methods for the detection of Listeria monocytogenes in cold-smoked salmon. Appl. Environ. Microbiol. 62, 822–824.
Anon. (2001) Protocol for the validation of alternative microbiological methods. NV-DOC.D-2001-04-25. Published by NordVal. Copenhagen, Denmark.
Bohnert, M.., Humbert, F. and Lombard, B. (2001) Validation of ISO microbiological methods. Detection of Salmonella according to draft standard pr EN ISO/ DIS 6579: 2000. Final Report. Ploufragan, France.
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Hoorfar, J., Cook, N. (2003). Critical Aspects of Standardization of PCR. In: Sachse, K., Frey, J. (eds) PCR Detection of Microbial Pathogens. Methods in Molecular Biology™, vol 216. Humana Press. https://doi.org/10.1385/1-59259-344-5:51
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DOI: https://doi.org/10.1385/1-59259-344-5:51
Publisher Name: Humana Press
Print ISBN: 978-1-58829-049-6
Online ISBN: 978-1-59259-344-6
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