Development of a Novel Peptide Nucleic Acid Probe for the Detection of Legionella spp. in Water Samples
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2022 |
| Outros Autores: | , , , , , , , |
| 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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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 |
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info:eu-repo/semantics/article |
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article |
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publishedVersion |
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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 |
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info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
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MDPI |
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MDPI |
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reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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