Abstract
Salmonella is still recognized as a major zoonotic pathogen for animals and humans (1). In many countries, it is the leading cause of all food-borne outbreaks and infections (2,3). Conventional cultural methods for the detection of Salmonella spp. in clinical human or animal material, food, feeding stuff or the environment require nonselective and selective incubation steps resulting in a 4 to 6 d isolation procedure (4,5). A more rapid detection of Salmonella spp. can be expected by the application of the polymerase chain reaction (PCR). Detection of Salmonella from food by PCR can be achieved within 24 h. Generally, the procedure includes (i) a pre-enrichment step in a nonselective medium to allow recovery of damaged cells and multiplication of the target organism; (ii) nucleic acid extraction from the culture; (iii) in vitro amplification of Salmonella spp. specific nucleic acid target DNA; and (iv) detection of the PCR product by gel electrophoresis and/or hybridization techniques. Several nonselective media for pre-enrichment have been developed for various food matrices, but buffered peptone water is most commonly used and recommended in the ISO 6579 standard on Microbiology of Food and Animal Feeding Stuffs-Horizontal Method for the Detection of Salmonella (6) as the medium of choice for pre-enrichment. Since many substances occurring in food can inhibit the thermostable DNA polymerase (7,8), the selection of a DNA extraction method depends on the food matrix.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Humphrey, T. ( 2000) Public-health aspects of Salmonella infection, in Salmonella in domestic animals (Way, C. and Way, A., ), CABI Publishing, Oxon, UK, pp. 245–263.
Tirado, C. and Schmidt, K. (2002) WHO Surveillance Programme for Control of Foodborne Infections and Intoxications: Results and trends across greater europe. J. Infect. 43, 80–84.
Wallace, D. J., Van Gilder, T., Shallow, S., et al. (2000) Incidence of food-borne illnesses reported by the food-borne diseases active surveillance network (FoodNet)-1997. FoodNet Working Group. J. Food Prot. 63, 807–809.
Waltman, W. D. ( 2000) Methods for the cultural isolation of Salmonella, in Salmonella in Domestic Animals (Wray, C. and Wray, A., ), CABI Publishing, Oxon, UK, pp. 355–372.
van der Zee, H. and Huis in’t Veld, J. H. J. (2000) Methods for rapid detection of Salmonella, in Salmonella in Domestic Animals (Way, C. and Way, A., ), CABI Publishing, Oxon, UK, pp. 373–391.
Anonymous (1993) Microbiology of food and animal feeding stuffs-Horizontal method for the detection of Salmonella (ISO 6579:1993). International Organization for Standardization.
Rossen, L., Nørskov, P., Holmstrøm, K., and Rasmussen, O. F. (1992) Inhibition of PCR by components of food samples, microbial diagnostic assays and DNA-extraction solutions. Int. J. Food Microbiol. 17, 37–45.
Wilson, I. G. (1997) Inhibition and facilitation of nucleic acid amplification. Appl. Environ. Microbiol. 63, 3741–3751.
Lantz, P.-G., Hahn-Hägerdal, B., and Rådström, P. (1994) Sample preparation methods in PCR-based detection of food pathogen. Trends Food Sci. Technol. 5, 384–389.
Lantz, P. G., al Soud, W. A., Knutsson, R., Hahn-Hagerdal, B., and Rådström, P. (2000) Biotechnical use of polymerase chain reaction for microbiological analysis of biological samples. Biotechnol. Annu. Rev. 5, 87–130.
Widjojoatmodjo, M. N., Fluit, A. C., Torensma, R., Keller, B. H., and Verhoef, J. (1991) Evaluation of the magnetic immuno PCR assay for rapid detection of Salmonella. Eur. J. Clin. Microbiol. Infect. Dis. 10, 935–938.
Rahn, K., De Grandis, S. A., Clarke, R. C., et al. (1992) Amplification of an invA gene sequence of Salmonella typhimurium by polymerase chain reaction as a specific method of detection of Salmonella. Mol. Cell. Probes 6, 271–279.
Aabo, S., Rasmussen, O. F., Rossen, L., Sorensen, P. D., and Olsen, J. E. (1993) Salmonella identification by the polymerase chain reaction. Mol. Cell. Probes 7, 171–178.
Doran, J. L., Collinson, S. K., Burian, J., et al. (1993) DNA-based diagnostic tests for Salmonella species targeting agfA, the structural gene for thin, aggregative fimbriae. J. Clin. Microbiol. 31, 2263–2273.
Jones, D. D., Law, R., and Bej, A. K. (1993) Detection of Salmonella spp. in oysters using polymerase chain reaction (PCR) and gene probes. J. Food. Sci. 58, 1191–1197.
Cohen, N. D., Neibergs, H. L., Wallis, D. E., Simpson, R. B., McGruder, E. D., and Hargis, B. M. (1994) Genus-specific detection of salmonellae in equine feces by use of the polymerase chain reaction. Am. J. Vet. Res. 55, 1049–1054.
Chen, S., Yee, A., Griffiths, M., et al. (1997) A rapid, sensitive and automated method for detection of Salmonella species in foods using AG-9600 AmpliSensor Analyzer. J. Appl. Microbiol. 83, 314–321.
Burkhalter, P. W., Müller, C., Luthy, J., and Candrian, U. (1995) Detection of Salmonella spp. in eggs: DNA analyses, culture techniques, and serology. J. AOACInt. 78, 1531–1537.
Kwang, J., Littledike, E. T., and Keen, J. E. (1996) Use of the polymerase chain reaction for Salmonella detection. Lett. Appl. Microbiol. 22, 46–51.
Bäumler, A. J., Heffron, F., and Reissbrodt, R. (1997) Rapid detection of Salmonella enterica with primers specific for iroB. J. Clin. Microbiol. 35, 1224–1230.
Makino, S., Kurazono, H., Chongsanguam, M., et al. (1999) Establishment of the PCR system specific to Salmonella spp. and its application for the inspection of food and fecal samples. J. Vet. Med. Sci. 61, 1245–1247.
Collazo, C. M. and Galán, J. E. (1997) The invasion-associated type-III protein secretion system in Salmonella-A review. Gene 192, 51–59.
Ausubel, F. M., Brent, R., Kingstone, R. E., et al. (1997) Current Protocols in Molecular Biology, John Wiley & Sons, Massachusetts, USA.
Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, CSH Laboratory Press, Cold Spring Harbor, New York, USA.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2003 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Malorny, B., Helmuth, R. (2003). Detection of Salmonella spp.. In: Sachse, K., Frey, J. (eds) PCR Detection of Microbial Pathogens. Methods in Molecular Biology™, vol 216. Humana Press. https://doi.org/10.1385/1-59259-344-5:275
Download citation
DOI: https://doi.org/10.1385/1-59259-344-5:275
Publisher Name: Humana Press
Print ISBN: 978-1-58829-049-6
Online ISBN: 978-1-59259-344-6
eBook Packages: Springer Protocols