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

2022-07-13Journal article Open access
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dc.contributor.authorNácher-Vázquez, Montserrat
dc.contributor.authorBarbosa, Ana
dc.contributor.authorArmelim, Inês
dc.contributor.authorAzevedo, Andreia Sofia
dc.contributor.authorAlmeida, Gonçalo Nieto
dc.contributor.authorPizarro, Cristina
dc.contributor.authorAzevedo, Nuno Filipe
dc.contributor.authorAlmeida, Carina
dc.contributor.authorCerqueira, Laura
dc.date.accessioned2023-03-21T10:19:42Z
dc.date.available2023-03-21T10:19:42Z
dc.date.issued2022-07-13
dc.descriptionThis article belongs to the Special Issue New Insights into Epidemiology, Detection and Characterization of Bacterial Pathogens.pt_PT
dc.description.abstractLegionella 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.pt_PT
dc.description.sponsorshipThis 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.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationMicroorganisms. 2022 Jul 13;10(7):1409. doi: 10.3390/microorganisms10071409pt_PT
dc.identifier.doi10.3390/microorganisms10071409pt_PT
dc.identifier.issn2076-2607
dc.identifier.urihttp://hdl.handle.net/10400.18/8583
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherMDPIpt_PT
dc.relationLaboratory for Process Engineering, Environment, Biotechnology and Energy
dc.relationLaboratory for Process Engineering, Environment, Biotechnology and Energy
dc.relation.publisherversionhttps://www.mdpi.com/2076-2607/10/7/1409pt_PT
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.subject16S rRNA Genept_PT
dc.subjectLegionella sp.pt_PT
dc.subjectFluorescence In Situ Hybridizationpt_PT
dc.subjectPeptide Nucleic Acid (PNA)pt_PT
dc.subjectWaterborne Detectionpt_PT
dc.subjectÁgua e Solopt_PT
dc.titleDevelopment of a Novel Peptide Nucleic Acid Probe for the Detection of Legionella spp. in Water Samplespt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleLaboratory for Process Engineering, Environment, Biotechnology and Energy
oaire.awardTitleLaboratory for Process Engineering, Environment, Biotechnology and Energy
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F00511%2F2020/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDP%2F00511%2F2020/PT
oaire.citation.issue7pt_PT
oaire.citation.startPage1409pt_PT
oaire.citation.titleMicroorganismspt_PT
oaire.citation.volume10pt_PT
oaire.fundingStream6817 - DCRRNI ID
oaire.fundingStream6817 - DCRRNI ID
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.embargofctAcesso de acordo com política editorial da revista.pt_PT
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
relation.isProjectOfPublication2f8cdc25-2875-4f26-8876-a08085c7f459
relation.isProjectOfPublication3210f5e0-5b59-47f4-b18d-c3b0fb4e5a93
relation.isProjectOfPublication.latestForDiscovery2f8cdc25-2875-4f26-8876-a08085c7f459

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