Simultaneous Screening of 24 Target Genes of Foodborne Pathogens in 35 Foodborne Outbreaks Using Multiplex Real-Time SYBR Green PCR Analysis

A set of 8 multiplex real-time SYBR Green PCR (SG-PCR) assays including 3 target primers and an internal amplification control (IAC) primer was simultaneously evaluated in 3 h or less with regard to detection of 24 target genes of 23 foodborne pathogens in 7 stool specimens of foodborne outbreak usi...

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Autores principales: Fukushima, Hiroshi, Kawase, Jun, Etoh, Yoshiki, Sugama, Kumiko, Yashiro, Shunshuke, Iida, Natsuko, Yamaguchi, Keiji
Formato: Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2010
Materias:
Research Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2948901/
https://www.ncbi.nlm.nih.gov/pubmed/20936159
http://dx.doi.org/10.1155/2010/864817
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author Fukushima, Hiroshi
Kawase, Jun
Etoh, Yoshiki
Sugama, Kumiko
Yashiro, Shunshuke
Iida, Natsuko
Yamaguchi, Keiji
author_facet Fukushima, Hiroshi
Kawase, Jun
Etoh, Yoshiki
Sugama, Kumiko
Yashiro, Shunshuke
Iida, Natsuko
Yamaguchi, Keiji
author_sort Fukushima, Hiroshi
collection PubMed
description A set of 8 multiplex real-time SYBR Green PCR (SG-PCR) assays including 3 target primers and an internal amplification control (IAC) primer was simultaneously evaluated in 3 h or less with regard to detection of 24 target genes of 23 foodborne pathogens in 7 stool specimens of foodborne outbreak using a 96-well reaction plate. This assay, combined with DNA extraction (QIAamp DNA Stool Mini kit), offered detection of greater than 10(3)-10(4) foodborne pathogens per g in stool specimens. The products formed were identified using melting point temperature (Tm) curve analysis. This assay was evaluated for the detection of foodborne pathogens in 33 out of 35 cases of foodborne outbreak, using 4 different PCR instruments in 5 different laboratories. No interference from the multiplex real-time SG-PCR assay, including IAC, was observed in stool specimens in any analysis. We found multiplex real-time SG-PCR assay for simultaneous detection of 24 target genes of foodborne pathogens to be comprehensive, rapid, inexpensive, accurate, of high selectivity, and good for screening probability.
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spelling pubmed-29489012010-10-08 Simultaneous Screening of 24 Target Genes of Foodborne Pathogens in 35 Foodborne Outbreaks Using Multiplex Real-Time SYBR Green PCR Analysis Fukushima, Hiroshi Kawase, Jun Etoh, Yoshiki Sugama, Kumiko Yashiro, Shunshuke Iida, Natsuko Yamaguchi, Keiji Int J Microbiol Research Article A set of 8 multiplex real-time SYBR Green PCR (SG-PCR) assays including 3 target primers and an internal amplification control (IAC) primer was simultaneously evaluated in 3 h or less with regard to detection of 24 target genes of 23 foodborne pathogens in 7 stool specimens of foodborne outbreak using a 96-well reaction plate. This assay, combined with DNA extraction (QIAamp DNA Stool Mini kit), offered detection of greater than 10(3)-10(4) foodborne pathogens per g in stool specimens. The products formed were identified using melting point temperature (Tm) curve analysis. This assay was evaluated for the detection of foodborne pathogens in 33 out of 35 cases of foodborne outbreak, using 4 different PCR instruments in 5 different laboratories. No interference from the multiplex real-time SG-PCR assay, including IAC, was observed in stool specimens in any analysis. We found multiplex real-time SG-PCR assay for simultaneous detection of 24 target genes of foodborne pathogens to be comprehensive, rapid, inexpensive, accurate, of high selectivity, and good for screening probability. Hindawi Publishing Corporation 2010 2010-09-28 /pmc/articles/PMC2948901/ /pubmed/20936159 http://dx.doi.org/10.1155/2010/864817 Text en Copyright © 2010 Hiroshi Fukushima 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
Fukushima, Hiroshi
Kawase, Jun
Etoh, Yoshiki
Sugama, Kumiko
Yashiro, Shunshuke
Iida, Natsuko
Yamaguchi, Keiji
Simultaneous Screening of 24 Target Genes of Foodborne Pathogens in 35 Foodborne Outbreaks Using Multiplex Real-Time SYBR Green PCR Analysis
title Simultaneous Screening of 24 Target Genes of Foodborne Pathogens in 35 Foodborne Outbreaks Using Multiplex Real-Time SYBR Green PCR Analysis
title_full Simultaneous Screening of 24 Target Genes of Foodborne Pathogens in 35 Foodborne Outbreaks Using Multiplex Real-Time SYBR Green PCR Analysis
title_fullStr Simultaneous Screening of 24 Target Genes of Foodborne Pathogens in 35 Foodborne Outbreaks Using Multiplex Real-Time SYBR Green PCR Analysis
title_full_unstemmed Simultaneous Screening of 24 Target Genes of Foodborne Pathogens in 35 Foodborne Outbreaks Using Multiplex Real-Time SYBR Green PCR Analysis
title_short Simultaneous Screening of 24 Target Genes of Foodborne Pathogens in 35 Foodborne Outbreaks Using Multiplex Real-Time SYBR Green PCR Analysis
title_sort simultaneous screening of 24 target genes of foodborne pathogens in 35 foodborne outbreaks using multiplex real-time sybr green pcr analysis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2948901/
https://www.ncbi.nlm.nih.gov/pubmed/20936159
http://dx.doi.org/10.1155/2010/864817
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