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Different patterns of azole resistance are linked to mutations in\n            <jats:italic>cyp51A<\/jats:italic>\n            . 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\n            <jats:named-content content-type=\"genus-species\">A. fumigatus<\/jats:named-content>\n            . PCRs targeting\n            <jats:italic>cyp51A<\/jats:italic>\n            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\n            <jats:named-content content-type=\"genus-species\">A. fumigatus<\/jats:named-content>\n            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 TR\n            <jats:sub>34<\/jats:sub>\n            , TR\n            <jats:sub>46<\/jats:sub>\n            , and TR\n            <jats:sub>53<\/jats:sub>\n            . Validation of the method was performed using a blind panel of 80\n            <jats:named-content content-type=\"genus-species\">A. fumigatus<\/jats:named-content>\n            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\n            <jats:named-content content-type=\"genus-species\">A. fumigatus<\/jats:named-content>\n            isolates and therefore facilitate selection of the best antifungal therapy in patients with aspergillosis.\n          <\/jats:p>","DOI":"10.1128\/aac.01083-17","type":"journal-article","created":{"date-parts":[[2017,9,12]],"date-time":"2017-09-12T00:40:28Z","timestamp":1505176828000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":18,"title":["Development and Validation of a High-Resolution Melting Assay To Detect Azole Resistance in Aspergillus fumigatus"],"prefix":"10.1128","volume":"61","author":[{"given":"L.","family":"Bernal-Mart\u00ednez","sequence":"first","affiliation":[{"name":"Mycology Reference Laboratory, National Centre for Microbiology, Instituto de Salud Carlos III, Madrid, Spain"},{"name":"Spanish Network for Research in Infectious Diseases, Instituto de Salud Carlos III, Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"H.","family":"Gil","sequence":"additional","affiliation":[{"name":"Mycology Reference Laboratory, National Centre for Microbiology, Instituto de Salud Carlos III, Madrid, Spain"},{"name":"European Program for Public Health Microbiology Training, European Centre for Disease Prevention and Control, Stockholm, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"O.","family":"Rivero-Men\u00e9ndez","sequence":"additional","affiliation":[{"name":"Mycology Reference Laboratory, National Centre for Microbiology, Instituto de Salud Carlos III, Madrid, Spain"},{"name":"Spanish Network for Research in Infectious Diseases, Instituto de Salud Carlos III, Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S.","family":"Gago","sequence":"additional","affiliation":[{"name":"Mycology Reference Laboratory, National Centre for Microbiology, Instituto de Salud Carlos III, Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M.","family":"Cuenca-Estrella","sequence":"additional","affiliation":[{"name":"Mycology Reference Laboratory, National Centre for Microbiology, Instituto de Salud Carlos III, Madrid, Spain"},{"name":"Spanish Network for Research in Infectious Diseases, Instituto de Salud Carlos III, Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"E.","family":"Mellado","sequence":"additional","affiliation":[{"name":"Mycology Reference Laboratory, National Centre for Microbiology, Instituto de Salud Carlos III, Madrid, Spain"},{"name":"Spanish Network for Research in Infectious Diseases, Instituto de Salud Carlos III, Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8651-4405","authenticated-orcid":false,"given":"A.","family":"Alastruey-Izquierdo","sequence":"additional","affiliation":[{"name":"Mycology Reference Laboratory, National Centre for Microbiology, Instituto de Salud Carlos III, Madrid, Spain"},{"name":"Spanish Network for Research in Infectious Diseases, Instituto de Salud Carlos III, Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","volume-title":"Risk assessment on the impact of environmental usage of triazoles on the development and spread of resistance to medical triazoles in Aspergillus species","author":"European Centre for Disease Prevention and Control","year":"2013","unstructured":"European Centre for Disease Prevention and Control. 2013. 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