Development of a Novel Peptide Nucleic Acid Probe for the Detection of Legionella spp. in Water Samples

Detalhes bibliográficos
Autor(a) principal: Nácher-Vázquez, Montserrat
Data de Publicação: 2022
Outros Autores: Barbosa, Ana, Armelim, Inês, Azevedo, Andreia Sofia, Almeida, Gonçalo Nieto, Pizarro, Cristina, Azevedo, Nuno Filipe, Almeida, Carina, Cerqueira, Laura
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10400.18/8583
Resumo: This article belongs to the Special Issue New Insights into Epidemiology, Detection and Characterization of Bacterial Pathogens.
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spelling Development of a Novel Peptide Nucleic Acid Probe for the Detection of Legionella spp. in Water Samples16S rRNA GeneLegionella sp.Fluorescence In Situ HybridizationPeptide Nucleic Acid (PNA)Waterborne DetectionÁgua e SoloThis article belongs to the Special Issue New Insights into Epidemiology, Detection and Characterization of Bacterial Pathogens.Legionella are opportunistic intracellular pathogens that are found throughout the environment. The Legionella contamination of water systems represents a serious social problem that can lead to severe diseases, which can manifest as both Pontiac fever and Legionnaires' disease (LD) infections. Fluorescence in situ hybridization using nucleic acid mimic probes (NAM-FISH) is a powerful and versatile technique for bacterial detection. By optimizing a peptide nucleic acid (PNA) sequence based on fluorescently selective binding to specific bacterial rRNA sequences, we established a new PNA-FISH method that has been successfully designed for the specific detection of the genus Legionella. The LEG22 PNA probe has shown great theoretical performance, presenting 99.9% specificity and 96.9% sensitivity. We also demonstrated that the PNA-FISH approach presents a good signal-to-noise ratio when applied in artificially contaminated water samples directly on filtration membranes or after cells elution. For water samples with higher turbidity (from cooling tower water systems), there is still the need for further method optimization in order to detect cellular contents and to overcome interferents' autofluorescence, which hinders probe signal visualization. Nevertheless, this work shows that the PNA-FISH approach could be a promising alternative for the rapid (3-4 h) and accurate detection of Legionella.This work was financially supported by: LA/P/0045/2020 (ALiCE), UIDB/00511/2020 and UIDP/00511/2020 (LEPABE), funded by national funds through the FCT/MCTES (PIDDAC); Projects POCI-01-0145-FEDER-029961, POCI-01-0145-FEDER-031011 and POCI-01-0145-FEDER-030431 funded by FEDER funds through COMPETE2020—Programa Operacional Competitividade e Internacionalização (POCI) and by national funds (PIDDAC) through FCT/MCTES. Montserrat Nácher-Vázquez and Laura Cerqueira were also financed by Project POCI-01-0145-FEDER-029961.MDPIRepositório Científico do Instituto Nacional de SaúdeNácher-Vázquez, MontserratBarbosa, AnaArmelim, InêsAzevedo, Andreia SofiaAlmeida, Gonçalo NietoPizarro, CristinaAzevedo, Nuno FilipeAlmeida, CarinaCerqueira, Laura2023-03-21T10:19:42Z2022-07-132022-07-13T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.18/8583engMicroorganisms. 2022 Jul 13;10(7):1409. doi: 10.3390/microorganisms100714092076-260710.3390/microorganisms10071409info: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:RCAAP2023-07-20T15:42:39Zoai:repositorio.insa.pt:10400.18/8583Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T18:43:13.677864Repositório 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 of a Novel Peptide Nucleic Acid Probe for the Detection of Legionella spp. in Water Samples
title Development of a Novel Peptide Nucleic Acid Probe for the Detection of Legionella spp. in Water Samples
spellingShingle Development of a Novel Peptide Nucleic Acid Probe for the Detection of Legionella spp. in Water Samples
Nácher-Vázquez, Montserrat
16S rRNA Gene
Legionella sp.
Fluorescence In Situ Hybridization
Peptide Nucleic Acid (PNA)
Waterborne Detection
Água e Solo
title_short Development of a Novel Peptide Nucleic Acid Probe for the Detection of Legionella spp. in Water Samples
title_full Development of a Novel Peptide Nucleic Acid Probe for the Detection of Legionella spp. in Water Samples
title_fullStr Development of a Novel Peptide Nucleic Acid Probe for the Detection of Legionella spp. in Water Samples
title_full_unstemmed Development of a Novel Peptide Nucleic Acid Probe for the Detection of Legionella spp. in Water Samples
title_sort Development of a Novel Peptide Nucleic Acid Probe for the Detection of Legionella spp. in Water Samples
author Nácher-Vázquez, Montserrat
author_facet Nácher-Vázquez, Montserrat
Barbosa, Ana
Armelim, Inês
Azevedo, Andreia Sofia
Almeida, Gonçalo Nieto
Pizarro, Cristina
Azevedo, Nuno Filipe
Almeida, Carina
Cerqueira, Laura
author_role author
author2 Barbosa, Ana
Armelim, Inês
Azevedo, Andreia Sofia
Almeida, Gonçalo Nieto
Pizarro, Cristina
Azevedo, Nuno Filipe
Almeida, Carina
Cerqueira, Laura
author2_role author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Repositório Científico do Instituto Nacional de Saúde
dc.contributor.author.fl_str_mv Nácher-Vázquez, Montserrat
Barbosa, Ana
Armelim, Inês
Azevedo, Andreia Sofia
Almeida, Gonçalo Nieto
Pizarro, Cristina
Azevedo, Nuno Filipe
Almeida, Carina
Cerqueira, Laura
dc.subject.por.fl_str_mv 16S rRNA Gene
Legionella sp.
Fluorescence In Situ Hybridization
Peptide Nucleic Acid (PNA)
Waterborne Detection
Água e Solo
topic 16S rRNA Gene
Legionella sp.
Fluorescence In Situ Hybridization
Peptide Nucleic Acid (PNA)
Waterborne Detection
Água e Solo
description This article belongs to the Special Issue New Insights into Epidemiology, Detection and Characterization of Bacterial Pathogens.
publishDate 2022
dc.date.none.fl_str_mv 2022-07-13
2022-07-13T00:00:00Z
2023-03-21T10:19:42Z
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/10400.18/8583
url http://hdl.handle.net/10400.18/8583
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Microorganisms. 2022 Jul 13;10(7):1409. doi: 10.3390/microorganisms10071409
2076-2607
10.3390/microorganisms10071409
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 MDPI
publisher.none.fl_str_mv MDPI
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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