Journal of Food, Agriculture and Environment




Vol 10, Issue 3&4,2012
Online ISSN: 1459-0263
Print ISSN: 1459-0255


Multiplex PCR detection of Salmonella spp., Listeria monocytogenesEscherichia coli O157:H7, and Staphylococcus aureus in processed foods using [TA10] simultaneous growth broth


Author(s):

Susumu Kawasaki 1*, Kaori Kusano 2, Ritsuko Arai 2, Toshihiro Komeda 2, Naoko Kamisaki-Horikoshi 3, Shinichi Kawamoto 1

Recieved Date: 2012-07-15, Accepted Date: 2012-09-24

Abstract:

A multiplex (m-)PCR method was developed for simultaneous detection of Salmonella spp., Listeria monocytogenes, Escherichia coli O157:H7 and Staphylococcus aureus in processed foods. The m-PCR detection protocol consisted of three steps: 1) 20-h pre-enrichment culturing, 2) simple DNA extraction, and 3) m-PCR detection. For pre-enrichment of pathogens from food samples, [TA10] simultaneous growth broth was used.  [TA10] broth does not contain selective agents and is of practical use for these four pathogenic bacteria without modification. For m-PCR detection, the targeted genes included a Salmonella species-specific sequence within a 1.8-kb HindIII DNA fragment, L. monocytogenes hlyA, E. coli O157:H7 eaeA, and S. aureus femA. The m-PCR assay with enrichment broth was evaluated in 13 types of spiked food samples (including pasteurized milk, yogurt, processed cheese, etc.), and targeted genes specific for each pathogen simultaneously. Assay sensitivity was 10° CFU for each pathogen in 25 g of sample with a 20-h enrichment period. This m-PCR method is reliable and useful for rapid screening of processed food samples contaminated with Salmonella spp., L. monocytogenes, E. coli O157:H7 or S. aureus

Keywords:

Multiplex PCR, detection, Salmonella spp., Listeria monocytogenes, Escherichia coli O157:H7, Staphylococcus aureus


Journal: Journal of Food, Agriculture and Environment
Year: 2012
Volume: 10
Issue: 3&4
Category: Food and Health
Pages: 261-266


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