Document Detail


In-house validation of a multiplex real-time PCR method for simultaneous detection of Salmonella spp., Escherichia coli O157 and Listeria monocytogenes.
MedLine Citation:
PMID:  23624537     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
A wide variety of qPCR methods currently exist for Salmonella spp., Escherichia coli O157 and Listeria monocytogenes detection. These methods target several genes and use different detection chemistries, either in simplex or in multiplex formats. However, the majority of these methods have not been carefully validated, and the number of validated methods that use multiplex qPCR is even lower. The aim of the present study was to develop and validate a multiplex qPCR method from previously validated simplex qPCR primers and probes. A modified broth medium was selected and primary and secondary enrichment times were further optimized. Efficiency of the newly combined qPCR system was comprised between 91% and 108%, for simplex and multiplex analyses. A total of 152 food and environmental, natural and spiked samples, were analyzed for the evaluation of the method obtaining values above 91% that were reached for all the quality parameters analyzed. A very low limit of detection (5cfu/25g after enrichment) for simultaneous identification of these 3 pathogens was obtained.
Authors:
Alejandro Garrido; María-José Chapela; Belén Román; Paula Fajardo; Juan M Vieites; Ana G Cabado
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2013-4-3
Journal Detail:
Title:  International journal of food microbiology     Volume:  164     ISSN:  1879-3460     ISO Abbreviation:  Int. J. Food Microbiol.     Publication Date:  2013 Apr 
Date Detail:
Created Date:  2013-4-29     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8412849     Medline TA:  Int J Food Microbiol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  92-98     Citation Subset:  -    
Copyright Information:
Copyright © 2013 Elsevier B.V. All rights reserved.
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