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United States Department of Agriculture

Agricultural Research Service

Research Project: MOLECULAR CHARACTERIZATION OF PATHOGENS AND THEIR RESPONSES TO ENVIRONMENTAL FACTORS

Location: Molecular Characterization of Foodborne Pathogens

Title: Development of a multiplex PCR detection kit for Salmonella spp., Listeria monocytogenes, and Escherichia coli O157:H7

Authors
item Kawasaki, Susumu -
item Fratamico, Pina
item Horikoshi, Naoko -
item Okada, Yukio -
item Takeshita, Kazuko -
item Sameshima, Takashi -
item Kawamoto, Shinichi -

Submitted to: Review Article
Publication Type: Review Article
Publication Acceptance Date: June 4, 2010
Publication Date: January 1, 2011
Citation: Kawasaki, S., Fratamico, P.M., Horikoshi, N., Okada, Y., Takeshita, K., Sameshima, T., Kawamoto, S. 2011. Development of a multiplex PCR detection kit for Salmonella spp., Listeria monocytogenes, and Escherichia coli O157:H7. Japan Agricultural Research Quarterly. 45:77-81.

Technical Abstract: A multiplex PCR method was developed for simultaneous detection of Salmonella spp., Listeria monocytogenes, and Escherichia coli O157:H7 in food samples, and the detection sensitivity for this method was 103 CFU/ml for each pathogen. When this method was used for the detection of each of pathogens in spiked pork samples, 1 cell per 25 g of inoculated sample could be detected within 24 h. Excellent agreement was obtained between the multiplex PCR assay and the conventional culture method, indicating that the multiplex PCR is a reliable and useful method for rapid screening of food products for Salmonella spp., L. monocytogenes, and E. coli O157:H7. These results prompted us to develop a multiplex PCR detection kit for quality control in the food industry. More than 50 types of spiked food products were tested with this kit at a commercial laboratory, and the detection rate of the PCR detection kit was higher than that of conventional culture methods. A detection sensitivity of 1 CFU per 25g of each pathogen was observed with this kit; therefore, this commercial kit provides a useful tool to ensure the safety of food tested by inspection laboratories and for the consumer.

Last Modified: 7/24/2014
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