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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...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Hindawi Publishing Corporation
2013
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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 |
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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 |
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