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A multiplex PCR assay for simultaneous detection of Escherichia coli O157:H7, Bacillus cereus, Vibrio parahaemolyticus, Salmonella spp., Listeria monocytogenes, and Staphylococcus aureus in Korean ready-to-eat food.
Foodborne Pathog Dis. 2014 Jul; 11(7):574-80.FP

Abstract

A multiplex polymerase chain reaction (PCR) assay was developed for simultaneous detection of Escherichia coli O157:H7, Bacillus cereus, Vibrio parahaemolyticus, Salmonella spp., Listeria monocytogenes, and Staphylococcus aureus in various Korean ready-to-eat foods. The six specific primer pairs for multiplex PCR were selected based on the O157 antigen (rfbE) gene of E. coli O157:H7, the DNA gyrase subunit B (gyrB) gene of B. cereus, the toxin regulatory protein (toxR) gene of V. parahaemolyticus, the invasion protein A (invA) gene of Salmonella spp., the hemolysin (hly) gene of L. monocytogenes, and the thermonuclease (nuc) gene of S. aureus. The 16S rRNA gene was targeted as an internal control gene in the presence of bacterial DNA. The specificity and sensitivity assays for multiplex primer pairs were investigated by testing different strains. When this multiplex PCR assay was applied to evaluate the validity of detecting six foodborne pathogens in artificially inoculated several ready-to-eat food samples, the assay was able to specifically simultaneously detect as few as 1 colony-forming unit/mL of each pathogen after enrichment for 12 h. Their presence in naturally contaminated samples also indicates that the developed multiplex PCR assay is an effective and informative supplement for practical use.

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Publisher Full Text (DOI)

Authors+Show Affiliations

Lee N
1 Food Safety Research Group, Korea Food Research Institute , Gyeonggi, Korea.
Kwon KY
No affiliation info available
Oh SK
No affiliation info available
Chang HJ
No affiliation info available
Chun HS
No affiliation info available
Choi SW
No affiliation info available

MeSH

Bacillus cereusColony Count, MicrobialDNA PrimersDNA, BacterialEscherichia coli O157Fast FoodsFood ContaminationFood MicrobiologyListeria monocytogenesMultiplex Polymerase Chain ReactionRNA, Ribosomal, 16SRepublic of KoreaSalmonellaSensitivity and SpecificityStaphylococcus aureusVibrio parahaemolyticus

Pub Type(s)

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

Language

eng

PubMed ID

24796416

Citation

Lee, Nari, et al. "A Multiplex PCR Assay for Simultaneous Detection of Escherichia Coli O157:H7, Bacillus Cereus, Vibrio Parahaemolyticus, Salmonella Spp., Listeria Monocytogenes, and Staphylococcus Aureus in Korean Ready-to-eat Food." Foodborne Pathogens and Disease, vol. 11, no. 7, 2014, pp. 574-80.
Lee N, Kwon KY, Oh SK, et al. A multiplex PCR assay for simultaneous detection of Escherichia coli O157:H7, Bacillus cereus, Vibrio parahaemolyticus, Salmonella spp., Listeria monocytogenes, and Staphylococcus aureus in Korean ready-to-eat food. Foodborne Pathog Dis. 2014;11(7):574-80.
Lee, N., Kwon, K. Y., Oh, S. K., Chang, H. J., Chun, H. S., & Choi, S. W. (2014). A multiplex PCR assay for simultaneous detection of Escherichia coli O157:H7, Bacillus cereus, Vibrio parahaemolyticus, Salmonella spp., Listeria monocytogenes, and Staphylococcus aureus in Korean ready-to-eat food. Foodborne Pathogens and Disease, 11(7), 574-80. https://doi.org/10.1089/fpd.2013.1638
Lee N, et al. A Multiplex PCR Assay for Simultaneous Detection of Escherichia Coli O157:H7, Bacillus Cereus, Vibrio Parahaemolyticus, Salmonella Spp., Listeria Monocytogenes, and Staphylococcus Aureus in Korean Ready-to-eat Food. Foodborne Pathog Dis. 2014;11(7):574-80. PubMed PMID: 24796416.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A multiplex PCR assay for simultaneous detection of Escherichia coli O157:H7, Bacillus cereus, Vibrio parahaemolyticus, Salmonella spp., Listeria monocytogenes, and Staphylococcus aureus in Korean ready-to-eat food. AU - Lee,Nari, AU - Kwon,Kyung Yoon, AU - Oh,Su Kyung, AU - Chang,Hyun-Joo, AU - Chun,Hyang Sook, AU - Choi,Sung-Wook, Y1 - 2014/05/05/ PY - 2014/5/7/entrez PY - 2014/5/7/pubmed PY - 2015/2/27/medline SP - 574 EP - 80 JF - Foodborne pathogens and disease JO - Foodborne Pathog Dis VL - 11 IS - 7 N2 - A multiplex polymerase chain reaction (PCR) assay was developed for simultaneous detection of Escherichia coli O157:H7, Bacillus cereus, Vibrio parahaemolyticus, Salmonella spp., Listeria monocytogenes, and Staphylococcus aureus in various Korean ready-to-eat foods. The six specific primer pairs for multiplex PCR were selected based on the O157 antigen (rfbE) gene of E. coli O157:H7, the DNA gyrase subunit B (gyrB) gene of B. cereus, the toxin regulatory protein (toxR) gene of V. parahaemolyticus, the invasion protein A (invA) gene of Salmonella spp., the hemolysin (hly) gene of L. monocytogenes, and the thermonuclease (nuc) gene of S. aureus. The 16S rRNA gene was targeted as an internal control gene in the presence of bacterial DNA. The specificity and sensitivity assays for multiplex primer pairs were investigated by testing different strains. When this multiplex PCR assay was applied to evaluate the validity of detecting six foodborne pathogens in artificially inoculated several ready-to-eat food samples, the assay was able to specifically simultaneously detect as few as 1 colony-forming unit/mL of each pathogen after enrichment for 12 h. Their presence in naturally contaminated samples also indicates that the developed multiplex PCR assay is an effective and informative supplement for practical use. SN - 1556-7125 UR - https://www.unboundmedicine.com/medline/citation/24796416/A_multiplex_PCR_assay_for_simultaneous_detection_of_Escherichia_coli_O157:H7_Bacillus_cereus_Vibrio_parahaemolyticus_Salmonella_spp__Listeria_monocytogenes_and_Staphylococcus_aureus_in_Korean_ready_to_eat_food_ DB - PRIME DP - Unbound Medicine ER -
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