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Multiplex real-time polymerase chain reaction assay for simultaneous detection and quantification of Salmonella species, Listeria monocytogenes, and Escherichia coli O157:H7 in ground pork samples.
Foodborne Pathog Dis. 2010 May; 7(5):549-54.FP

Abstract

Salmonella sp., Listeria monocytogenes, and Escherichia coli O157:H7 are foodborne pathogens capable of causing serious gastrointestinal illness. We previously described simultaneous detection of these pathogens by multiplex polymerase chain reaction (PCR) in 44 types of spiked food samples, including meat, produce, fish, and dairy products, targeting genes specific for each pathogen. Based on the previous work, a multiplex real-time PCR assay using fluorescent probes was developed to detect and accurately quantify Salmonella sp., L. monocytogenes, and E. coli O157:H7 in ground pork samples. The detection sensitivity for this method was 2.0 x 10(2) CFU/mL for each pathogen, and the quantification range was 10(2)-10(7) CFU/mL with a high correlation coefficient (R(2) > 0.99) and high PCR efficiency (84.2% to 99.2%). When this protocol was used for the detection of each of the pathogens in spiked pork samples, one cell per 25 g of inoculated sample after enrichment for 20 h could be detected within 24 h. As a result, this multiplex real-time PCR assay will be valuable as a screening method for foods contaminated with these pathogens.

Authors+Show Affiliations

Food Hygiene Laboratory, National Food Research Institute, Tsukuba, Japan. skawasa@affrc.go.jpNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20132032

Citation

Kawasaki, Susumu, et al. "Multiplex Real-time Polymerase Chain Reaction Assay for Simultaneous Detection and Quantification of Salmonella Species, Listeria Monocytogenes, and Escherichia Coli O157:H7 in Ground Pork Samples." Foodborne Pathogens and Disease, vol. 7, no. 5, 2010, pp. 549-54.
Kawasaki S, Fratamico PM, Horikoshi N, et al. Multiplex real-time polymerase chain reaction assay for simultaneous detection and quantification of Salmonella species, Listeria monocytogenes, and Escherichia coli O157:H7 in ground pork samples. Foodborne Pathog Dis. 2010;7(5):549-54.
Kawasaki, S., Fratamico, P. M., Horikoshi, N., Okada, Y., Takeshita, K., Sameshima, T., & Kawamoto, S. (2010). Multiplex real-time polymerase chain reaction assay for simultaneous detection and quantification of Salmonella species, Listeria monocytogenes, and Escherichia coli O157:H7 in ground pork samples. Foodborne Pathogens and Disease, 7(5), 549-54. https://doi.org/10.1089/fpd.2009.0465
Kawasaki S, et al. Multiplex Real-time Polymerase Chain Reaction Assay for Simultaneous Detection and Quantification of Salmonella Species, Listeria Monocytogenes, and Escherichia Coli O157:H7 in Ground Pork Samples. Foodborne Pathog Dis. 2010;7(5):549-54. PubMed PMID: 20132032.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Multiplex real-time polymerase chain reaction assay for simultaneous detection and quantification of Salmonella species, Listeria monocytogenes, and Escherichia coli O157:H7 in ground pork samples. AU - Kawasaki,Susumu, AU - Fratamico,Pina M, AU - Horikoshi,Naoko, AU - Okada,Yukio, AU - Takeshita,Kazuko, AU - Sameshima,Takashi, AU - Kawamoto,Shinichi, PY - 2010/2/6/entrez PY - 2010/2/6/pubmed PY - 2010/8/21/medline SP - 549 EP - 54 JF - Foodborne pathogens and disease JO - Foodborne Pathog Dis VL - 7 IS - 5 N2 - Salmonella sp., Listeria monocytogenes, and Escherichia coli O157:H7 are foodborne pathogens capable of causing serious gastrointestinal illness. We previously described simultaneous detection of these pathogens by multiplex polymerase chain reaction (PCR) in 44 types of spiked food samples, including meat, produce, fish, and dairy products, targeting genes specific for each pathogen. Based on the previous work, a multiplex real-time PCR assay using fluorescent probes was developed to detect and accurately quantify Salmonella sp., L. monocytogenes, and E. coli O157:H7 in ground pork samples. The detection sensitivity for this method was 2.0 x 10(2) CFU/mL for each pathogen, and the quantification range was 10(2)-10(7) CFU/mL with a high correlation coefficient (R(2) > 0.99) and high PCR efficiency (84.2% to 99.2%). When this protocol was used for the detection of each of the pathogens in spiked pork samples, one cell per 25 g of inoculated sample after enrichment for 20 h could be detected within 24 h. As a result, this multiplex real-time PCR assay will be valuable as a screening method for foods contaminated with these pathogens. SN - 1556-7125 UR - https://www.unboundmedicine.com/medline/citation/20132032/Multiplex_real_time_polymerase_chain_reaction_assay_for_simultaneous_detection_and_quantification_of_Salmonella_species_Listeria_monocytogenes_and_Escherichia_coli_O157:H7_in_ground_pork_samples_ DB - PRIME DP - Unbound Medicine ER -