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BioMed Research International
Volume 2013 (2013), Article ID 295050, 10 pages
http://dx.doi.org/10.1155/2013/295050
Research Article

A New Protocol to Detect Multiple Foodborne Pathogens with PCR Dipstick DNA Chromatography after a Six-Hour Enrichment Culture in a Broad-Range Food Pathogen Enrichment Broth

1Division of Anaerobe Research, Life Science Research Center, Gifu University, 1-1 Yanagido, Gifu 501-1194, Japan
2Department of Microbiology, Gifu University Graduate School of Medicine, 1-1 Yanagido, Gifu 501-1194, Japan
3Division of Food Hygiene, Department of Animal and Food Hygiene, Obihiro University of Agriculture & Veterinary Medicine Inada-cho, Obihiro, Hokkaido 080-8555, Japan
4Department of Domestic Science, Kyoto Seibo College, Kyoto 612-0878, Japan

Received 27 September 2013; Revised 5 November 2013; Accepted 5 November 2013

Academic Editor: Hiroshi Asakura

Copyright © 2013 Masahiro Hayashi et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

A quick foodborne pathogen screening method after six-hour enrichment culture with a broad-range food pathogen enrichment broth is described. Pathogenic factors of Salmonella enterica, Shigella spp., enteroinvasive Escherichia coli, and enterohemorrhagic E. coli are amplified with a cocktail primer and rapid polymerase chain reaction (PCR), which finishes amplification in 30 min. The PCR amplicon was differentiated with a dipstick DNA chromatography assay in 5–10 min. Starting from a four- to six-hour enrichment culture, this assay was finished within 45 min. Detection sensitivity of this protocol was less than 2.5 CFU/25 g for S. enterica and 3.3 CFU/25 g for enterohemorrhagic E. coli in spiked ground meat experiments.