Utilize este identificador para referenciar este registo:
https://hdl.handle.net/1822/55190
Registo completo
Campo DC | Valor | Idioma |
---|---|---|
dc.contributor.author | Rocha, Rui | por |
dc.contributor.author | Almeida, Carina | por |
dc.contributor.author | Azevedo, Nuno F. | por |
dc.date.accessioned | 2018-06-29T10:13:52Z | - |
dc.date.available | 2018-06-29T10:13:52Z | - |
dc.date.issued | 2018-05 | - |
dc.identifier.citation | Rocha, Rui; Almeida, Carina; Azevedo, Nuno F., Influence of the fixation/permeabilization step on peptide nucleic acid fluorescence in situ hybridization (PNA-FISH) for the detection of bacteria. PLoS One, 13(5), e0196522, 2018 | por |
dc.identifier.issn | 1932-6203 | por |
dc.identifier.uri | https://hdl.handle.net/1822/55190 | - |
dc.description.abstract | Fluorescence in situ Hybridization (FISH) is a versatile, widespread and widely- used technique in microbiology. The first step of FISHfixation/permeabilizationis crucial to the outcome of the method. This work aimed to systematically evaluate fixation/permeabilization protocols employing ethanol, triton X-100 and lysozyme in conjugation with paraformaldehyde for Peptide Nucleic Acid (PNA)-FISH. Response surface methodology was used to optimize these protocols for Gram-negative (Escherichia coli and Pseudomonas fluorescens) and Gram-positive species (Listeria innocua, Staphylococcus epidermidis and Bacillus cereus). In general, the optimal PNA-FISH fluorescent outcome in Gram-positive bacteria was obtained employing harsher permeabilization conditions when compared to Gram-negative optimal protocols. The observed differences arise from the intrinsic cell envelope properties of each species and the ability of the fixation/permeabilization compounds to effectively increase the permeability of these structures while maintaining structural integrity. Ultimately, the combination of paraformaldehyde and ethanol proved to have significantly superior performance for all tested bacteria, especially for Gram-positive species (p<0.05). | por |
dc.description.sponsorship | This work was the result of the projects: POCI-01-0145-FEDER-006939 (Laboratory for Process Engineering, Environment, Biotechnology and Energy - UID/EQU/00511/2013) and POCI-010145-FEDER-006684 (Centerof Biological Engineering - UID/EQU/00511/2013)funded by the European Regional Development Fund (ERDF), through COMPETE2020- Programa Operacional Competitividade e Internacionalização (POCI) and by national funds, throughFCT - Fundação para a Ciênciae a Tecnologia. NORTE-01-0145-FEDER000005 - LEPABE-2-ECO-INNOVATIONand NORTE-01-0145-FEDER-000004- BioTecNorte operation,supported by North Portugal Regional Operational Programme (NORTE 2020), under the Portugal 2020 PartnershipAgreement, throughthe EuropeanRegional Development Fund (ERDF). PhD Fellowship SFRH/BDE/51910/2012 supported by national funds through FCT - Fundação para a Ciênciae a Tecnologia. ENMed/0003/2014 NanoDiaBac and PTDC/DTP-PIC/4562/2014 Coded-FISHfunded by national funds throughFCT - Fundac ¸ão para a Ciênciae a Tecnologia. Rui Rocha works for and receives salary from Biomode S.A, a companythat developsand commercializes in vitro diagnostickits for microorganism detection, using PNA-FISHtechnology. Also, Carina Almeida and Nuno F. Azevedo are foundersand shareholders of Biomode SA. The specific roles of these authors are articulated in the ‘author contributions’ section. The funders had no role in study design, data collectionand analysis, decision to publish, or preparation of the manuscript. | por |
dc.language.iso | eng | por |
dc.publisher | Public Library of Science | por |
dc.relation | info:eu-repo/grantAgreement/FCT/5876/147284/PT | por |
dc.relation | info:eu-repo/grantAgreement/FCT/5876/147284/PT | por |
dc.relation | info:eu-repo/grantAgreement/FCT/SFRH/SFRH%2FBDE%2F51910%2F2012/PT | por |
dc.relation | info:eu-repo/grantAgreement/FCT/3599-PPCDT/138727/PT | por |
dc.relation | PTDC/DTP-PIC/4562/2014 | por |
dc.rights | openAccess | por |
dc.subject | Ethanol fixation | por |
dc.subject | Gram positive bacteria | por |
dc.subject | Bacillus cereus | por |
dc.subject | Gram negative bacteria | por |
dc.subject | Staphylococcus epidermidis | por |
dc.subject | Fluorescent in situ hybridization | por |
dc.subject | Pseudomonas fluorescens | por |
dc.subject | Cell hybridization | por |
dc.title | Influence of the fixation/permeabilization step on peptide nucleic acid fluorescence in situ hybridization (PNA-FISH) for the detection of bacteria | por |
dc.type | article | - |
dc.peerreviewed | yes | por |
dc.relation.publisherversion | http://journals.plos.org/plosone/ | por |
dc.comments | CEB47740 | por |
oaire.citationStartPage | e0196522 | por |
oaire.citationIssue | 5 | por |
oaire.citationConferencePlace | United States | - |
oaire.citationVolume | 13 | por |
dc.date.updated | 2018-06-02T11:19:26Z | - |
dc.identifier.doi | 10.1371/journal.pone.0196522 | por |
dc.identifier.pmid | 29851961 | por |
dc.description.publicationversion | info:eu-repo/semantics/publishedVersion | por |
dc.subject.wos | Science & Technology | por |
sdum.journal | PLoS One | por |
Aparece nas coleções: | CEB - Publicações em Revistas/Séries Internacionais / Publications in International Journals/Series |
Ficheiros deste registo:
Ficheiro | Descrição | Tamanho | Formato | |
---|---|---|---|---|
document_47740_1.pdf | 3,2 MB | Adobe PDF | Ver/Abrir |