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dc.contributor.author | Bernal-Martinez, Leticia | |
dc.contributor.author | Gil, Horacio | |
dc.contributor.author | Rivero-Menendez, Olga | |
dc.contributor.author | Gago Prieto, Sara | |
dc.contributor.author | Cuenca-Estrella, Manuel | |
dc.contributor.author | Mellado, Emilia | |
dc.contributor.author | Alastruey-Izquierdo, Ana | |
dc.date.accessioned | 2020-06-11T09:08:22Z | |
dc.date.available | 2020-06-11T09:08:22Z | |
dc.date.issued | 2017 | |
dc.identifier.citation | Antimicrob Agents Chemother . 2017 Nov 22;61(12):e01083-17. | es_ES |
dc.identifier.uri | http://hdl.handle.net/20.500.12105/10338 | |
dc.description.abstract | The global emergence of azole-resistant Aspergillus fumigatus strains is a growing public health concern. Different patterns of azole resistance are linked to mutations in cyp51A Therefore, accurate characterization of the mechanisms underlying azole resistance is critical to guide selection of the most appropriate antifungal agent for patients with aspergillosis. This study describes a new sequencing-free molecular screening tool for early detection of the most frequent mutations known to be associated with azole resistance in A. fumigatus PCRs targeting cyp51A mutations at positions G54, Y121, G448, and M220 and targeting different tandem repeats (TRs) in the promoter region were designed. All PCRs were performed simultaneously, using the same cycling conditions. Amplicons were then distinguished using a high-resolution melting assay. For standardization, 30 well-characterized azole-resistant A. fumigatus strains were used, yielding melting curve clusters for different resistance mechanisms for each target and allowing detection of the most frequent azole resistance mutations, i.e., G54E, G54V, G54R, G54W, Y121F, M220V, M220I, M220T, M220K, and G448S, and the tandem repeats TR34, TR46, and TR53 Validation of the method was performed using a blind panel of 80 A. fumigatus azole-susceptible or azole-resistant strains. All strains included in the blind panel were properly classified as susceptible or resistant with the developed method. The implementation of this screening method can reduce the time needed for the detection of azole-resistant A. fumigatus isolates and therefore facilitate selection of the best antifungal therapy in patients with aspergillosis. | es_ES |
dc.description.sponsorship | This study was financed by a research project from the Fondo de Investigación Sanitaria (FIS) (project PI13/02145). It was supported by Plan Nacional de I+D+i and Instituto de Salud Carlos III, Subdirección General de Redes y Centros de Investigación Cooperativa, Ministerio de Economía y Competitividad, and the Spanish Network for Research in Infectious Diseases (REIPI grant RD12/0015), cofinanced by the European Development Regional Fund. L.B.-M. has a research contract from the Spanish Network for Research in Infectious Diseases (REIPI grant RD12/0015). O.R.-M. holds a predoctoral fellowship from the Fondo de Investigaciones Sanitarias (grant FI14CIII/00025). S.G. was supported by a research fellowship from the Fondo de Investigaciones Sanitarias of the Spanish Ministry of Science and Innovation (grant FI10/00464). E.M. was supported by a project from the Spanish Fondo de Investigación Sanitaria (FIS grant PI15CIII/00019). | es_ES |
dc.language.iso | eng | es_ES |
dc.publisher | American Society for Microbiology (ASM) | es_ES |
dc.type.hasVersion | AM | es_ES |
dc.rights.uri | http://creativecommons.org/licenses/by-nc-sa/4.0/ | * |
dc.subject.mesh | Antifungal Agents | es_ES |
dc.subject.mesh | Aspergillosis | es_ES |
dc.subject.mesh | Aspergillus fumigatus | es_ES |
dc.subject.mesh | Cytochrome P-450 Enzyme System | es_ES |
dc.subject.mesh | Drug Resistance, Fungal | es_ES |
dc.subject.mesh | Fungal Proteins | es_ES |
dc.subject.mesh | Humans | es_ES |
dc.subject.mesh | Microbial Sensitivity Tests | es_ES |
dc.subject.mesh | Nucleic Acid Denaturation | es_ES |
dc.subject.mesh | Polymerase Chain Reaction | es_ES |
dc.subject.mesh | Promoter Regions, Genetic | es_ES |
dc.subject.mesh | Tandem Repeat Sequences | es_ES |
dc.title | Development and Validation of a High-Resolution Melting Assay To Detect Azole Resistance in Aspergillus fumigatus. | es_ES |
dc.type | journal article | es_ES |
dc.rights.license | Atribución-NoComercial-CompartirIgual 4.0 Internacional | * |
dc.identifier.pubmedID | 28893791 | es_ES |
dc.format.volume | 61 | es_ES |
dc.format.number | 12 | es_ES |
dc.identifier.doi | 10.1128/AAC.01083-17 | es_ES |
dc.contributor.funder | Instituto de Salud Carlos III | |
dc.contributor.funder | RETICS-Investigación en Patología Infecciosa (REIPI-ISCIII) (España) | |
dc.contributor.funder | Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF) | |
dc.description.peerreviewed | Sí | es_ES |
dc.identifier.e-issn | 1098-6596 | es_ES |
dc.relation.publisherversion | https://doi.org/10.1128/AAC.01083-17 | es_ES |
dc.identifier.journal | Antimicrobial agents and chemotherapy | es_ES |
dc.repisalud.centro | ISCIII::Centro Nacional de Microbiología | es_ES |
dc.repisalud.institucion | ISCIII | es_ES |
dc.relation.projectID | info:eu_repo/grantAgreement/ES/PI13/02145 | es_ES |
dc.relation.projectID | info:eu_repo/grantAgreement/ES/RD12/0015 | es_ES |
dc.relation.projectID | info:eu_repo/grantAgreement/ES/FI14CIII/00025 | es_ES |
dc.relation.projectID | info:eu_repo/grantAgreement/ES/FI10/00464 | es_ES |
dc.relation.projectID | info:eu_repo/grantAgreement/ES/PI15CIII/00019 | es_ES |
dc.rights.accessRights | open access | es_ES |