title: 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.
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.
authors.affiliation: Division of Anaerobe Research, Life Science Research Center, Gifu University, 1-1 Yanagido, Gifu 501-1194, Japan ; Department of Microbiology, Gifu University Graduate School of Medicine, 1-1 Yanagido, Gifu 501-1194, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Hayashi
authors.affiliation: Department of Microbiology, Gifu University Graduate School of Medicine, 1-1 Yanagido, Gifu 501-1194, Japan.
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Natori
authors.affiliation: Department of Microbiology, Gifu University Graduate School of Medicine, 1-1 Yanagido, Gifu 501-1194, Japan.
authors.forename: Sayoko
authors.initials: S
authors.lastname: Kubota-Hayashi
authors.affiliation: Department of Microbiology, Gifu University Graduate School of Medicine, 1-1 Yanagido, Gifu 501-1194, Japan.
authors.forename: Machiko
authors.initials: M
authors.lastname: Miyata
authors.affiliation: Department of Microbiology, Gifu University Graduate School of Medicine, 1-1 Yanagido, Gifu 501-1194, Japan.
authors.forename: Kiyofumi
authors.initials: K
authors.lastname: Ohkusu
authors.affiliation: Division of Food Hygiene, Department of Animal and Food Hygiene, Obihiro University of Agriculture & Veterinary Medicine Inada-cho, Obihiro, Hokkaido 080-8555, Japan.
authors.forename: Keiko
authors.initials: K
authors.lastname: Kawamoto
authors.affiliation: Division of Food Hygiene, Department of Animal and Food Hygiene, Obihiro University of Agriculture & Veterinary Medicine Inada-cho, Obihiro, Hokkaido 080-8555, Japan.
authors.forename: Hisao
authors.initials: H
authors.lastname: Kurazono
authors.affiliation: Department of Domestic Science, Kyoto Seibo College, Kyoto 612-0878, Japan.
authors.forename: Souichi
authors.initials: S
authors.lastname: Makino
authors.affiliation: Department of Microbiology, Gifu University Graduate School of Medicine, 1-1 Yanagido, Gifu 501-1194, Japan.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Ezaki
mesh_terms: D015169:Colony Count, Microbial; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity; D005516:Food Microbiology; D005517:Foodborne Diseases / Q000235:genetics* / Q000382:microbiology; D006801:Humans; D016133:Polymerase Chain Reaction; D019779:Salmonella enterica / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity; D012760:Shigella / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity
chemical_list: D004269:DNA, Bacterial