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A multiplex real-time polymerase chain reaction for simultaneous detection of Salmonella spp., Escherichia coli O157, and Listeria monocytogenes in meat products.
Foodborne Pathog Dis. 2010 Jun; 7(6):619-28.FP

Abstract

To achieve an effective detection of Salmonella spp., Escherichia coli O157, and Listeria monocytogenes in meat products, a multiplex real-time polymerase chain reaction (PCR) coupled with a multipathogen enrichment strategy was developed in this study. Pathogen-specific DNA sequences in the invA, rfbE, and hlyA genes were employed to design primers and TaqMan probes for identifying Salmonella spp., E. coli O157, and L. monocytogenes, respectively. An internal amplification control (IAC) utilizing a novel DNA sequence from human adenovirus was incorporated into the multiplex PCR assay to indicate false-negative results. Concurrent amplifications of multiple targets and IAC were thoroughly evaluated and optimized to minimize PCR competitions. Combined with a multipathogen enrichment in a selective enrichment broth for Salmonella, Escherichia, and Listeria (SEL), the multiplex real-time PCR assay was able to simultaneously detect all of the three organisms in artificially contaminated ground beef at a detection sensitivity of <18 CFU/10 g ground beef. Applying the assay to 26 retail meat samples including beef, chicken, turkey, and pork revealed that 12 samples were positive for one of the organisms and 3 samples were positive for two of the organisms after a 20-h enrichment in SEL. The remaining meat samples tested negative for all of the organisms by only showing amplification of the IAC. These results were confirmed by traditional culture methods testing for each individual species. Taken together, the multiplex real-time PCR assay combined with multipathogen enrichment is a rapid and reliable method for effectively screening single or multiple pathogen occurrences in various meat products.

Authors+Show Affiliations

Joint Sino-U.S. Food Safety Research Center and Bor Luh Food Safety Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

20113204

Citation

Suo, Biao, et al. "A Multiplex Real-time Polymerase Chain Reaction for Simultaneous Detection of Salmonella Spp., Escherichia Coli O157, and Listeria Monocytogenes in Meat Products." Foodborne Pathogens and Disease, vol. 7, no. 6, 2010, pp. 619-28.
Suo B, He Y, Tu SI, et al. A multiplex real-time polymerase chain reaction for simultaneous detection of Salmonella spp., Escherichia coli O157, and Listeria monocytogenes in meat products. Foodborne Pathog Dis. 2010;7(6):619-28.
Suo, B., He, Y., Tu, S. I., & Shi, X. (2010). A multiplex real-time polymerase chain reaction for simultaneous detection of Salmonella spp., Escherichia coli O157, and Listeria monocytogenes in meat products. Foodborne Pathogens and Disease, 7(6), 619-28. https://doi.org/10.1089/fpd.2009.0430
Suo B, et al. A Multiplex Real-time Polymerase Chain Reaction for Simultaneous Detection of Salmonella Spp., Escherichia Coli O157, and Listeria Monocytogenes in Meat Products. Foodborne Pathog Dis. 2010;7(6):619-28. PubMed PMID: 20113204.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A multiplex real-time polymerase chain reaction for simultaneous detection of Salmonella spp., Escherichia coli O157, and Listeria monocytogenes in meat products. AU - Suo,Biao, AU - He,Yiping, AU - Tu,Shu-I, AU - Shi,Xianming, PY - 2010/2/2/entrez PY - 2010/2/2/pubmed PY - 2010/9/2/medline SP - 619 EP - 28 JF - Foodborne pathogens and disease JO - Foodborne Pathog Dis VL - 7 IS - 6 N2 - To achieve an effective detection of Salmonella spp., Escherichia coli O157, and Listeria monocytogenes in meat products, a multiplex real-time polymerase chain reaction (PCR) coupled with a multipathogen enrichment strategy was developed in this study. Pathogen-specific DNA sequences in the invA, rfbE, and hlyA genes were employed to design primers and TaqMan probes for identifying Salmonella spp., E. coli O157, and L. monocytogenes, respectively. An internal amplification control (IAC) utilizing a novel DNA sequence from human adenovirus was incorporated into the multiplex PCR assay to indicate false-negative results. Concurrent amplifications of multiple targets and IAC were thoroughly evaluated and optimized to minimize PCR competitions. Combined with a multipathogen enrichment in a selective enrichment broth for Salmonella, Escherichia, and Listeria (SEL), the multiplex real-time PCR assay was able to simultaneously detect all of the three organisms in artificially contaminated ground beef at a detection sensitivity of <18 CFU/10 g ground beef. Applying the assay to 26 retail meat samples including beef, chicken, turkey, and pork revealed that 12 samples were positive for one of the organisms and 3 samples were positive for two of the organisms after a 20-h enrichment in SEL. The remaining meat samples tested negative for all of the organisms by only showing amplification of the IAC. These results were confirmed by traditional culture methods testing for each individual species. Taken together, the multiplex real-time PCR assay combined with multipathogen enrichment is a rapid and reliable method for effectively screening single or multiple pathogen occurrences in various meat products. SN - 1556-7125 UR - https://www.unboundmedicine.com/medline/citation/20113204/A_multiplex_real_time_polymerase_chain_reaction_for_simultaneous_detection_of_Salmonella_spp__Escherichia_coli_O157_and_Listeria_monocytogenes_in_meat_products_ L2 - https://www.liebertpub.com/doi/10.1089/fpd.2009.0430?url_ver=Z39.88-2003&amp;rfr_id=ori:rid:crossref.org&amp;rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -