Cargando…

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

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Hayashi, Masahiro, Natori, Tatsuya, Kubota-Hayashi, Sayoko, Miyata, Machiko, Ohkusu, Kiyofumi, Kawamoto, Keiko, Kurazono, Hisao, Makino, Souichi, Ezaki, Takayuki
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3865639/
https://www.ncbi.nlm.nih.gov/pubmed/24364031
http://dx.doi.org/10.1155/2013/295050
_version_ 1782296058855948288
author Hayashi, Masahiro
Natori, Tatsuya
Kubota-Hayashi, Sayoko
Miyata, Machiko
Ohkusu, Kiyofumi
Kawamoto, Keiko
Kurazono, Hisao
Makino, Souichi
Ezaki, Takayuki
author_facet Hayashi, Masahiro
Natori, Tatsuya
Kubota-Hayashi, Sayoko
Miyata, Machiko
Ohkusu, Kiyofumi
Kawamoto, Keiko
Kurazono, Hisao
Makino, Souichi
Ezaki, Takayuki
author_sort Hayashi, Masahiro
collection PubMed
description 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.
format Online
Article
Text
id pubmed-3865639
institution National Center for Biotechnology Information
language English
publishDate 2013
publisher Hindawi Publishing Corporation
record_format MEDLINE/PubMed
spelling pubmed-38656392013-12-22 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 Hayashi, Masahiro Natori, Tatsuya Kubota-Hayashi, Sayoko Miyata, Machiko Ohkusu, Kiyofumi Kawamoto, Keiko Kurazono, Hisao Makino, Souichi Ezaki, Takayuki Biomed Res Int Research Article 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. Hindawi Publishing Corporation 2013 2013-12-02 /pmc/articles/PMC3865639/ /pubmed/24364031 http://dx.doi.org/10.1155/2013/295050 Text en Copyright © 2013 Masahiro Hayashi et al. https://creativecommons.org/licenses/by/3.0/ 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.
spellingShingle Research Article
Hayashi, Masahiro
Natori, Tatsuya
Kubota-Hayashi, Sayoko
Miyata, Machiko
Ohkusu, Kiyofumi
Kawamoto, Keiko
Kurazono, Hisao
Makino, Souichi
Ezaki, Takayuki
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
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
title_full 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
title_fullStr 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
title_full_unstemmed 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
title_short 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
title_sort 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
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3865639/
https://www.ncbi.nlm.nih.gov/pubmed/24364031
http://dx.doi.org/10.1155/2013/295050
work_keys_str_mv AT hayashimasahiro anewprotocoltodetectmultiplefoodbornepathogenswithpcrdipstickdnachromatographyafterasixhourenrichmentcultureinabroadrangefoodpathogenenrichmentbroth
AT natoritatsuya anewprotocoltodetectmultiplefoodbornepathogenswithpcrdipstickdnachromatographyafterasixhourenrichmentcultureinabroadrangefoodpathogenenrichmentbroth
AT kubotahayashisayoko anewprotocoltodetectmultiplefoodbornepathogenswithpcrdipstickdnachromatographyafterasixhourenrichmentcultureinabroadrangefoodpathogenenrichmentbroth
AT miyatamachiko anewprotocoltodetectmultiplefoodbornepathogenswithpcrdipstickdnachromatographyafterasixhourenrichmentcultureinabroadrangefoodpathogenenrichmentbroth
AT ohkusukiyofumi anewprotocoltodetectmultiplefoodbornepathogenswithpcrdipstickdnachromatographyafterasixhourenrichmentcultureinabroadrangefoodpathogenenrichmentbroth
AT kawamotokeiko anewprotocoltodetectmultiplefoodbornepathogenswithpcrdipstickdnachromatographyafterasixhourenrichmentcultureinabroadrangefoodpathogenenrichmentbroth
AT kurazonohisao anewprotocoltodetectmultiplefoodbornepathogenswithpcrdipstickdnachromatographyafterasixhourenrichmentcultureinabroadrangefoodpathogenenrichmentbroth
AT makinosouichi anewprotocoltodetectmultiplefoodbornepathogenswithpcrdipstickdnachromatographyafterasixhourenrichmentcultureinabroadrangefoodpathogenenrichmentbroth
AT ezakitakayuki anewprotocoltodetectmultiplefoodbornepathogenswithpcrdipstickdnachromatographyafterasixhourenrichmentcultureinabroadrangefoodpathogenenrichmentbroth
AT hayashimasahiro newprotocoltodetectmultiplefoodbornepathogenswithpcrdipstickdnachromatographyafterasixhourenrichmentcultureinabroadrangefoodpathogenenrichmentbroth
AT natoritatsuya newprotocoltodetectmultiplefoodbornepathogenswithpcrdipstickdnachromatographyafterasixhourenrichmentcultureinabroadrangefoodpathogenenrichmentbroth
AT kubotahayashisayoko newprotocoltodetectmultiplefoodbornepathogenswithpcrdipstickdnachromatographyafterasixhourenrichmentcultureinabroadrangefoodpathogenenrichmentbroth
AT miyatamachiko newprotocoltodetectmultiplefoodbornepathogenswithpcrdipstickdnachromatographyafterasixhourenrichmentcultureinabroadrangefoodpathogenenrichmentbroth
AT ohkusukiyofumi newprotocoltodetectmultiplefoodbornepathogenswithpcrdipstickdnachromatographyafterasixhourenrichmentcultureinabroadrangefoodpathogenenrichmentbroth
AT kawamotokeiko newprotocoltodetectmultiplefoodbornepathogenswithpcrdipstickdnachromatographyafterasixhourenrichmentcultureinabroadrangefoodpathogenenrichmentbroth
AT kurazonohisao newprotocoltodetectmultiplefoodbornepathogenswithpcrdipstickdnachromatographyafterasixhourenrichmentcultureinabroadrangefoodpathogenenrichmentbroth
AT makinosouichi newprotocoltodetectmultiplefoodbornepathogenswithpcrdipstickdnachromatographyafterasixhourenrichmentcultureinabroadrangefoodpathogenenrichmentbroth
AT ezakitakayuki newprotocoltodetectmultiplefoodbornepathogenswithpcrdipstickdnachromatographyafterasixhourenrichmentcultureinabroadrangefoodpathogenenrichmentbroth