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A Novel Multiplex PCR Assay for Rapid and Simultaneous Detection of Five Pathogenic Bacteria: Escherichia coli O157:H7, Salmonella, Staphylococcus aureus, Listeria monocytogenes, and Vibrio parahaemolyticus

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Abstract:

Rapid and sensitive detection techniques for foodborne pathogens are important to the food industry. However, traditional detection methods rely on bacterial culture in combination with biochemical tests, a process that typically takes 4 to 7 days to complete. Thus, this study was conducted to address the issue of time lag inherent in traditional methods by developing a novel PCR assay for each of five foodborne pathogenic bacteria. This new system consists of a simultaneous screening method using multiplex PCR in a single reaction tube for the rapid and sensitive detection of each of the five bacteria. Specific primers for multiplex PCR amplification of the Shiga-like toxin (verotoxin type II), femA (cytoplasmic protein), toxR (transmembrane DNA binding protein), iap (invasive associative protein), and invA (invasion protein A) genes were designed to allow simultaneous detection of Escherichia coli O157:H7, Staphylococcus aureus, Vibrio parahaemolyticus, Listeria monocytogenes, and Salmonella, respectively. To confirm the specificity of each primer pair for the respective target gene, three types of experiments were carried out using boiled cell lysates and their DNAs. In the multiplex PCR with mixed DNA samples, specific bands for corresponding genes were simultaneously detected from a single reaction. The detection of all five foodborne pathogenic bacteria could be completed in less than 24 h with this novel PCR method.

Document Type: Research Article

Affiliations: 1: Samsung Everland Inc., Food Research & Development Center, 50 Mabuk-dong, Giheung-gu, Yongin 446-912, South Korea; Department of Genetic Engineering, Sungkyunkwan University, 300 Chunchon-dong, Jangan-gu, Suwon 440-746, South Korea 2: Samsung Everland Inc., Food Research & Development Center, 50 Mabuk-dong, Giheung-gu, Yongin 446-912, South Korea 3: Kyungin Regional Korea Food & Drug Administration, Hazard Analysis Laboratory, #120, Juan-1-dong, Nam-gu, Incheon, South Korea 4: Korea Food & Drug Administration, Food Microbiology Division, 231 Jinheungno, Eunpyeong-gu, Seoul, South Korea 5: KoGene BioTech., Inc., R&D Center, 510 Chuksun Hyundae Bldg., Chuksun-dong 80, Chongro-gu, Seoul, South Korea 6: Department of Genetic Engineering, Sungkyunkwan University, 300 Chunchon-dong, Jangan-gu, Suwon 440-746, South Korea

Publication date: July 1, 2007

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    First published in 1937, the Journal of Food Protection®, is a refereed monthly publication. Each issue contains scientific research and authoritative review articles reporting on a variety of topics in food science pertaining to food safety and quality. The Journal is internationally recognized as the leading publication in the field of food microbiology with a readership exceeding 11,000 scientists from 70 countries. The Journal of Food Protection® is indexed in Index Medicus, Current Contents, BIOSIS, PubMed, Medline, and many others.

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