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Multiplex PCR assay for the detection and quantification of Campylobacter spp., Escherichia coli O157:H7, and Salmonella serotypes in water samples.
MedLine Citation:
PMID:  21204924     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Three pathogens, Campylobacter, Salmonella, and Shiga-toxin-producing Escherichia coli, are leading causes of bacterial gastroenteritis in the United States and worldwide. Although these three bacteria are typically considered food-borne pathogens, outbreaks have been reported due to contaminated drinking water and irrigation water. The aim of this research was to develop two types of PCR assays that could detect and quantify three pathogens, Campylobacter spp., E. coli O157:H7, and Salmonella spp., in watershed samples. In conventional PCR, three target strains were detected by multiplex PCR (m-PCR) using each specific primer pair simultaneously. Under optimized m-PCR conditions, the assay produced a 90-bp product for Campylobacter jejuni, a 150-bp product for E. coli O157:H7, and a 262-bp product for Salmonella Typhimurium, and the limitation of detection was approximately 700 copies for all three bacteria. In addition, real-time PCR was performed to quantify the three pathogens using SYBR green fluorescence. The assay was designed so that each target had a different melting temperature [C. jejuni (80.1 °C), E. coli O157:H7 (83.3 °C), and S. Typhimurium (85.9 °C)]. Therefore, this system could quantify and distinguish three pathogens simultaneously in a single reaction.
Authors:
Si Hong Park; Irene Hanning; Robin Jarquin; Philip Moore; Dan J Donoghue; Ann M Donoghue; Steven C Ricke
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Publication Detail:
Type:  Journal Article     Date:  2011-01-14
Journal Detail:
Title:  FEMS microbiology letters     Volume:  316     ISSN:  1574-6968     ISO Abbreviation:  FEMS Microbiol. Lett.     Publication Date:  2011 Mar 
Date Detail:
Created Date:  2011-02-01     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7705721     Medline TA:  FEMS Microbiol Lett     Country:  England    
Other Details:
Languages:  eng     Pagination:  7-15     Citation Subset:  IM    
Copyright Information:
© 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.
Affiliation:
Cell and Molecular Biology Program, University of Arkansas, Fayetteville, AR, USA Center for Food Safety-IFSE and Department of Food Science, University of Arkansas, Fayetteville, AR, USA Department of Poultry Science, University of Arkansas, Fayetteville, AR, USA Poultry Production and Poultry Safety Research Unit, Agricultural Research Services, USDA, Fayetteville, AR, USA.
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