2026-06-14T03:45:06Zhttps://repisalud.isciii.es/rest/oai/requestoai:repisalud.isciii.es:20.500.12105/103382024-10-31T11:52:57Zcom_20.500.12105_2052com_20.500.12105_2051col_20.500.12105_19609
Repisalud
author
Bernal-Martinez, Leticia
author
Gil, Horacio
author
Rivero-Menendez, Olga
author
Gago Prieto, Sara
author
Cuenca-Estrella, Manuel
author
Mellado, Emilia
author
Alastruey-Izquierdo, Ana
funder
Instituto de Salud Carlos III
funder
RETICS-Investigación en Patología Infecciosa (REIPI-ISCIII) (España)
funder
Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF)
2020-06-11T09:08:22Z
2020-06-11T09:08:22Z
2017
Antimicrob Agents Chemother . 2017 Nov 22;61(12):e01083-17.
10.1128/AAC.01083-17
1098-6596
Antimicrobial agents and chemotherapy
28893791
http://hdl.handle.net/20.500.12105/10338
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.
eng
Development and Validation of a High-Resolution Melting Assay To Detect Azole Resistance in Aspergillus fumigatus.
journal article
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URL
https://repisalud.isciii.es/bitstreams/ad118e2b-4f1f-4e3b-a497-fd7561c8a309/download
File
MD5
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application/pdf
DevelopmentAndValidationOfaAHigh_2017.pdf
URL
https://repisalud.isciii.es/bitstreams/9e8e0e41-5d5a-4ff5-90d1-2af1d37316a6/download
File
MD5
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text/plain
DevelopmentAndValidationOfaAHigh_2017.pdf.txt