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We analyzed 17 Aspergillus isolates from respiratory specimens of 15 critically ill patients with COVID-19 in S\u00e3o Paulo State, Brazil, during 2020\u20132021. Patients were retrospectively classified according to ECMM\/ISHAM criteria as probable CAPA (n = 5), possible CAPA (n = 7), or non-CAPA (n = 3). Complementary reassessment using the 2024 FUNDICU definitions classified patients as probable invasive pulmonary aspergillosis (IPA; n = 3), probable tracheobronchial aspergillosis (TBA; n = 1), or as not meeting these definitions (n = 9); two were ineligible due to classical EORTC\/MSGERC host factors (n = 2). Species identification was performed by benA sequencing and MALDI-TOF MS. Susceptibility to amphotericin B (AMB), itraconazole (ITZ), voriconazole, and posaconazole (PCZ) was determined according to CLSI M38-A2, and\u00a0cyp51A and its promoter were analyzed in A. fumigatus. The predominant species were A. fumigatus (n = 7) and A. flavus (n = 5), followed by A. tamarii (n = 2), A. parasiticus, A. terreus, and A. welwitschiae (n = 1 each). All A. fumigatus isolates exhibited non-wild-type phenotypes (NWT) against PCZ, with some also showing reduced susceptibility to ITZ and AMB. Silent cyp51A mutations occurred in three A. fumigatus isolates, while several section Flavi isolates displayed multiazole NWT. These findings highlight diagnostic limitations and emerging azole resistance, supporting susceptibility testing and surveillance.<\/jats:p>","DOI":"10.1093\/mmy\/myag092","type":"journal-article","created":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T11:52:42Z","timestamp":1788954762000},"source":"Crossref","is-referenced-by-count":0,"title":["Molecular characterization and antifungal susceptibility of\n                    <i>Aspergillus<\/i>\n                    spp. isolated from critically ill COVID-19 patients: Implications for suspected COVID-19-associated pulmonary aspergillosis (CAPA) in a Brazilian tertiary referral hospital"],"prefix":"10.1093","volume":"64","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3306-3118","authenticated-orcid":false,"given":"Tiago Alexandre","family":"Cocio","sequence":"first","affiliation":[{"name":"Ribeir\u00e3o Preto Medical School, University of S\u00e3o Paulo (USP) Department of Internal Medicine, , Avenida Bandeirantes, 3900\u2014Monte Alegre, Ribeir\u00e3o Preto, SP 14049-900 ,","place":["Brazil"]},{"name":"Laboratory of Medical Investigation in Mycology (LIM53), Institute of Tropical Medicine, Faculty of Medicine, Department of Dermatology, University of S\u00e3o Paulo (USP) , Avenida Dr. En\u00e9as de Carvalho Aguiar, 470\u2014Cerqueira C\u00e9sar, S\u00e3o Paulo, SP 05403-000 ,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vivian Caso","family":"Coelho","sequence":"additional","affiliation":[{"name":"Laboratory of Medical Investigation in Immunology (LIM 48), Hospital das Cl\u00ednicas, Faculty of Medicine, University of S\u00e3o Paulo (USP) , Avenida Dr. En\u00e9as de Carvalho Aguiar, 255\u2014 Cerqueira C\u00e9sar, S\u00e3o Paulo, SP 05403-000 ,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ludmilla","family":"Tonani","sequence":"additional","affiliation":[{"name":"Toxicology and Food Science, School of Pharmaceutical Sciences of Ribeir\u00e3o Preto, University of S\u00e3o Paulo (USP) Department of Clinical Analyses, , Avenida do Caf\u00e9, s\/n\u2014 Monte Alegre, Ribeir\u00e3o Preto, SP 14040-903 ,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ingrid Gon\u00e7alves","family":"Costa\u00a0\u2013\u00a0Leite","sequence":"additional","affiliation":[{"name":"Laboratory of Medical Investigation in Mycology (LIM53), Institute of Tropical Medicine, Faculty of Medicine, Department of Dermatology, University of S\u00e3o Paulo (USP) , Avenida Dr. En\u00e9as de Carvalho Aguiar, 470\u2014Cerqueira C\u00e9sar, S\u00e3o Paulo, SP 05403-000 ,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marcia Regina von Zeska","family":"Kress","sequence":"additional","affiliation":[{"name":"Toxicology and Food Science, School of Pharmaceutical Sciences of Ribeir\u00e3o Preto, University of S\u00e3o Paulo (USP) Department of Clinical Analyses, , Avenida do Caf\u00e9, s\/n\u2014 Monte Alegre, Ribeir\u00e3o Preto, SP 14040-903 ,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4322-312X","authenticated-orcid":false,"given":"Gilda Maria Barbaro","family":"Del Negro","sequence":"additional","affiliation":[{"name":"Laboratory of Medical Investigation in Mycology (LIM53), Institute of Tropical Medicine, Faculty of Medicine, Department of Dermatology, University of S\u00e3o Paulo (USP) , Avenida Dr. En\u00e9as de Carvalho Aguiar, 470\u2014Cerqueira C\u00e9sar, S\u00e3o Paulo, SP 05403-000 ,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1335-8808","authenticated-orcid":false,"given":"Marcia de Souza Carvalho","family":"Melhem","sequence":"additional","affiliation":[{"name":"Laboratory of Medical Investigation in Mycology (LIM53), Institute of Tropical Medicine, Faculty of Medicine, Department of Dermatology, University of S\u00e3o Paulo (USP) , Avenida Dr. En\u00e9as de Carvalho Aguiar, 470\u2014Cerqueira C\u00e9sar, S\u00e3o Paulo, SP 05403-000 ,","place":["Brazil"]},{"name":"School of Medicine, Federal University of Mato Grosso do Sul (UFMS) , Avenida Costa e Silva, s\/n\u2014Bairro Universit\u00e1rio, Campo Grande, MS 79070-900 ,","place":["Brazil"]},{"name":"School of Medicine, S\u00e3o Paulo State University (UNESP), Graduate Program of Tropical Diseases , Distrito de Rubi\u00e3o J\u00fanior, s\/n, Botucatu, SP 18618-687 ,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C\u00e9lia","family":"Soares","sequence":"additional","affiliation":[{"name":"CEB-Biological Engineering Center, Micoteca da Universidade do Minho, Universidade do Minho , Campus de Gualtar, 4710-057 Braga ,","place":["Portugal"]},{"name":"LABBELS (Associate Laboratory, Braga\/Guimar\u00e3es), University of Minho , Campus de Gualtar, 4710-057 Braga 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