Development and application of Peptide Nucleic Acid Fluorescence in situ Hybridization for the specific detection of Listeria monocytogenes

Detalhes bibliográficos
Autor(a) principal: Rocha, Rui
Data de Publicação: 2019
Outros Autores: Sousa, J., Cerqueira, Laura, Vieira, Maria João, Almeida, Carina, Azevedo, Nuno Filipe Ribeiro Pinto Oliveira
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/1822/57495
Resumo: Supplementary data to this article can be found online at https:// doi.org/10.1016/j.fm.2018.12.009.
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spelling Development and application of Peptide Nucleic Acid Fluorescence in situ Hybridization for the specific detection of Listeria monocytogenesListeria monocytogenesPNA-FISHBlocker probeEnrichment procedureScience & TechnologySupplementary data to this article can be found online at https:// doi.org/10.1016/j.fm.2018.12.009.Listeria monocytogenes is one of the most important foodborne pathogens due to the high hospitalization and mortality rates associated to an outbreak. Several new molecular methods that accelerate the identification of L. monocytogenes have been developed, however conventional culture-based methods still remain the gold standard. In this work we developed a novel Peptide Nucleic Acid Fluorescence in situ Hybridization (PNA-FISH) method for the specific detection of L. monocytogenes. The method was based on an already existing PNA probe, LmPNA1253, coupled with a novel blocker probe in a 1:2 ratio. The method was optimized for the detection of L. monocytogenes in food samples through an evaluation of several rich and selective enrichment broths. The best outcome was achieved using One Broth Listeria in a two-step enrichment of 24h plus 18h. For validation in food samples, ground beef, ground pork, milk, lettuce and cooked shrimp were artificially contaminated with two ranges of inoculum: a low level (0.22CFU/25g or mL) and a high level (210CFU/25g or mL). The PNA-FISH method performed well in all types of food matrices, presenting an overall accuracy of 99% and a detection limit of 0.5CFU/25g or mL of food sample.Nuno R. da Silva acknowledges the financial support byFundac¸ ão para a Ciência e Tecnologia (FCT) of the Portuguese’s Min-istry for Science, Technology and Higher Education (MCTES), in theframework of the POCH – Programa Operacional Capital Humano,co-funded by the European Social Fund (SFRH/BD/111825/2015).This work was also supported by FCT under the scope ofthe strategic funding of UID/BIO/04469 unit and COMPETE2020 (POCI-01-0145-FEDER-006684) and BioTecNorte opera-tion (NORTE-01-0145-FEDER-000004) funded by the EuropeanRegional Development Fund under the scope of Norte2020 – Pro-grama Operacional Regional do Norte.info:eu-repo/semantics/publishedVersionElsevierUniversidade do MinhoRocha, RuiSousa, J.Cerqueira, LauraVieira, Maria JoãoAlmeida, CarinaAzevedo, Nuno Filipe Ribeiro Pinto Oliveira2019-062019-06-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/1822/57495engRocha, Rui; Sousa, J.; Cerqueira, Laura; Vieira, Maria João; Almeida, Carina; Azevedo, Nuno F., Development and application of Peptide Nucleic Acid Fluorescence in situ Hybridization for the specific detection of Listeria monocytogenes. Food Microbiology, 80, 1-8, 20190740-00201095-999810.1016/j.fm.2018.12.00930704592http://www.elsevier.com/locate/issn/07400020info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-21T12:41:18Zoai:repositorium.sdum.uminho.pt:1822/57495Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:38:16.038301Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Development and application of Peptide Nucleic Acid Fluorescence in situ Hybridization for the specific detection of Listeria monocytogenes
title Development and application of Peptide Nucleic Acid Fluorescence in situ Hybridization for the specific detection of Listeria monocytogenes
spellingShingle Development and application of Peptide Nucleic Acid Fluorescence in situ Hybridization for the specific detection of Listeria monocytogenes
Rocha, Rui
Listeria monocytogenes
PNA-FISH
Blocker probe
Enrichment procedure
Science & Technology
title_short Development and application of Peptide Nucleic Acid Fluorescence in situ Hybridization for the specific detection of Listeria monocytogenes
title_full Development and application of Peptide Nucleic Acid Fluorescence in situ Hybridization for the specific detection of Listeria monocytogenes
title_fullStr Development and application of Peptide Nucleic Acid Fluorescence in situ Hybridization for the specific detection of Listeria monocytogenes
title_full_unstemmed Development and application of Peptide Nucleic Acid Fluorescence in situ Hybridization for the specific detection of Listeria monocytogenes
title_sort Development and application of Peptide Nucleic Acid Fluorescence in situ Hybridization for the specific detection of Listeria monocytogenes
author Rocha, Rui
author_facet Rocha, Rui
Sousa, J.
Cerqueira, Laura
Vieira, Maria João
Almeida, Carina
Azevedo, Nuno Filipe Ribeiro Pinto Oliveira
author_role author
author2 Sousa, J.
Cerqueira, Laura
Vieira, Maria João
Almeida, Carina
Azevedo, Nuno Filipe Ribeiro Pinto Oliveira
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade do Minho
dc.contributor.author.fl_str_mv Rocha, Rui
Sousa, J.
Cerqueira, Laura
Vieira, Maria João
Almeida, Carina
Azevedo, Nuno Filipe Ribeiro Pinto Oliveira
dc.subject.por.fl_str_mv Listeria monocytogenes
PNA-FISH
Blocker probe
Enrichment procedure
Science & Technology
topic Listeria monocytogenes
PNA-FISH
Blocker probe
Enrichment procedure
Science & Technology
description Supplementary data to this article can be found online at https:// doi.org/10.1016/j.fm.2018.12.009.
publishDate 2019
dc.date.none.fl_str_mv 2019-06
2019-06-01T00:00:00Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/1822/57495
url http://hdl.handle.net/1822/57495
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Rocha, Rui; Sousa, J.; Cerqueira, Laura; Vieira, Maria João; Almeida, Carina; Azevedo, Nuno F., Development and application of Peptide Nucleic Acid Fluorescence in situ Hybridization for the specific detection of Listeria monocytogenes. Food Microbiology, 80, 1-8, 2019
0740-0020
1095-9998
10.1016/j.fm.2018.12.009
30704592
http://www.elsevier.com/locate/issn/07400020
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
institution RCAAP
reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
repository.mail.fl_str_mv
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