Simultaneous and Sensitive Detection of Three Foodborne Pathogens by Multiplex PCR, Capillary Gel Electrophoresis, and Laser-Induced Fluorescence

The simultaneous detection of Staphylococcus aureus, Listeria monocytogenes, and Salmonella spp. has been approached by a new multiplex PCR-based procedure followed by capillary gel electrophoresis with laser-induced fluorescence detection (multiplex-PCR-CGE-LIF). As compared to slab gel electrophor...

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Detalhes bibliográficos
Autores: Alarcón, Benito, García-Cañas, Virginia, Cifuentes, Alejandro, González García, Ramón, Aznar, Rosa
Tipo de documento: artigo
Data de publicação:2004
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositório:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/12120
Acesso em linha:http://hdl.handle.net/10261/12120
Access Level:Acceso aberto
Palavra-chave:Multiplex PCR
CGE
LIF
Food analysis
foodborne pathogens
DNA analysis
Salmonella spp.
Staphylococcus aureus
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spelling Simultaneous and Sensitive Detection of Three Foodborne Pathogens by Multiplex PCR, Capillary Gel Electrophoresis, and Laser-Induced FluorescenceAlarcón, BenitoGarcía-Cañas, VirginiaCifuentes, AlejandroGonzález García, RamónAznar, RosaMultiplex PCRCGELIFFood analysisfoodborne pathogensDNA analysisSalmonella spp.Staphylococcus aureusThe simultaneous detection of Staphylococcus aureus, Listeria monocytogenes, and Salmonella spp. has been approached by a new multiplex PCR-based procedure followed by capillary gel electrophoresis with laser-induced fluorescence detection (multiplex-PCR-CGE-LIF). As compared to slab gel electrophoresis, the use of CGE-LIF improved from 10- to 1000-fold the sensitivity of the multiplex PCR analysis, allowing the detection of 2.6 × 103 cfu mL-1 of S. aureus, 570 cfu mL-1 of L. monocytogenes, and 790 cfu mL-1 of Salmonella in artificially inoculated food, without enrichment. Following 6 h of enrichment, as low as 260, 79, and 57 cfu mL-1 of S. aureus, L. monocytogenes, and Salmonella, respectively, were detected. The CGE-LIF method is shown to be reproducible, providing relative standard deviation (RSD) values lower than 0.8% for analysis time and lower than 5.8% for peak areas. The multiplex-PCR-CGE-LIF proved a powerful analytical tool to detect various food pathogens simultaneously in a fast, reproducible, and sensitive way.Peer reviewedAmerican Chemical Society200920092004info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_650188 bytestext/plainhttp://hdl.handle.net/10261/12120reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Ingléshttp://pubs.acs.org/doi/pdf/10.1021/jf049038binfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/121202026-05-22T06:33:51Z
dc.title.none.fl_str_mv Simultaneous and Sensitive Detection of Three Foodborne Pathogens by Multiplex PCR, Capillary Gel Electrophoresis, and Laser-Induced Fluorescence
title Simultaneous and Sensitive Detection of Three Foodborne Pathogens by Multiplex PCR, Capillary Gel Electrophoresis, and Laser-Induced Fluorescence
spellingShingle Simultaneous and Sensitive Detection of Three Foodborne Pathogens by Multiplex PCR, Capillary Gel Electrophoresis, and Laser-Induced Fluorescence
Alarcón, Benito
Multiplex PCR
CGE
LIF
Food analysis
foodborne pathogens
DNA analysis
Salmonella spp.
Staphylococcus aureus
title_short Simultaneous and Sensitive Detection of Three Foodborne Pathogens by Multiplex PCR, Capillary Gel Electrophoresis, and Laser-Induced Fluorescence
title_full Simultaneous and Sensitive Detection of Three Foodborne Pathogens by Multiplex PCR, Capillary Gel Electrophoresis, and Laser-Induced Fluorescence
title_fullStr Simultaneous and Sensitive Detection of Three Foodborne Pathogens by Multiplex PCR, Capillary Gel Electrophoresis, and Laser-Induced Fluorescence
title_full_unstemmed Simultaneous and Sensitive Detection of Three Foodborne Pathogens by Multiplex PCR, Capillary Gel Electrophoresis, and Laser-Induced Fluorescence
title_sort Simultaneous and Sensitive Detection of Three Foodborne Pathogens by Multiplex PCR, Capillary Gel Electrophoresis, and Laser-Induced Fluorescence
dc.creator.none.fl_str_mv Alarcón, Benito
García-Cañas, Virginia
Cifuentes, Alejandro
González García, Ramón
Aznar, Rosa
author Alarcón, Benito
author_facet Alarcón, Benito
García-Cañas, Virginia
Cifuentes, Alejandro
González García, Ramón
Aznar, Rosa
author_role author
author2 García-Cañas, Virginia
Cifuentes, Alejandro
González García, Ramón
Aznar, Rosa
author2_role author
author
author
author
dc.subject.none.fl_str_mv Multiplex PCR
CGE
LIF
Food analysis
foodborne pathogens
DNA analysis
Salmonella spp.
Staphylococcus aureus
topic Multiplex PCR
CGE
LIF
Food analysis
foodborne pathogens
DNA analysis
Salmonella spp.
Staphylococcus aureus
description The simultaneous detection of Staphylococcus aureus, Listeria monocytogenes, and Salmonella spp. has been approached by a new multiplex PCR-based procedure followed by capillary gel electrophoresis with laser-induced fluorescence detection (multiplex-PCR-CGE-LIF). As compared to slab gel electrophoresis, the use of CGE-LIF improved from 10- to 1000-fold the sensitivity of the multiplex PCR analysis, allowing the detection of 2.6 × 103 cfu mL-1 of S. aureus, 570 cfu mL-1 of L. monocytogenes, and 790 cfu mL-1 of Salmonella in artificially inoculated food, without enrichment. Following 6 h of enrichment, as low as 260, 79, and 57 cfu mL-1 of S. aureus, L. monocytogenes, and Salmonella, respectively, were detected. The CGE-LIF method is shown to be reproducible, providing relative standard deviation (RSD) values lower than 0.8% for analysis time and lower than 5.8% for peak areas. The multiplex-PCR-CGE-LIF proved a powerful analytical tool to detect various food pathogens simultaneously in a fast, reproducible, and sensitive way.
publishDate 2004
dc.date.none.fl_str_mv 2004
2009
2009
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/12120
url http://hdl.handle.net/10261/12120
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv http://pubs.acs.org/doi/pdf/10.1021/jf049038b
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 88 bytes
text/plain
dc.publisher.none.fl_str_mv American Chemical Society
publisher.none.fl_str_mv American Chemical Society
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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