{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,2,21]],"date-time":"2025-02-21T09:27:49Z","timestamp":1740130069956,"version":"3.37.3"},"reference-count":14,"publisher":"Oxford University Press (OUP)","issue":"5","license":[{"start":{"date-parts":[[2022,2,23]],"date-time":"2022-02-23T00:00:00Z","timestamp":1645574400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2022,4,27]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:sec>\n                  <jats:title>Objectives<\/jats:title>\n                  <jats:p>Current reference susceptibility testing methods of Aspergillus require visual reading, which is subjective and necessitates experienced staff. We compared spectrophotometric and visual MIC reading of EUCAST E.Def 9.3.2 susceptibility testing of Aspergillus fumigatus for a large collection of isolates with different azole resistance mechanisms.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>A. fumigatus (n = 200) were examined, including 62 WT and 138 non-WT with the following alterations: TR34\/L98H (n = 57), TR46\/Y121F\/T289A (n = 54) or single point mutations (n = 27). EUCAST E.Def 9.3.2 susceptibility testing was performed for amphotericin B, itraconazole, voriconazole, posaconazole and isavuconazole. MICs were determined after 48\u2005h of incubation visually and spectrophotometrically, as the lowest concentration corresponding to a 1%, 3%, 5%, 10% or 15% OD increase above the background OD. The best spectrophotometric endpoint (SPE) was identified based on the highest essential agreement (EA; \u00b11 two-fold dilution) and categorical agreement (CA) and fewer very major errors (VMEs) and major errors (MEs).<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>\u03a4he best SPEs were 5% and 10% for all drugs. The best agreement between visual and spectrophotometric MICs was found with the 10% growth endpoint, which resulted in identical median MICs with 90% of differences being \u22641 two-fold and higher EA (91%\u2013100%) and CA (100%) and no VMEs and MEs compared with the 5% endpoint (77%\u2013100%, 96%\u201398%, 0% and 0%\u20134%, respectively).<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>Spectrophotometric MIC reading can be used for A. fumigatus susceptibility testing and for detecting azole resistance. A visual inspection of the plate should be performed to confirm equal inoculation, absence of well contamination and proper growth, and to identify potential uncommon phenotypes or subpopulations.<\/jats:p>\n               <\/jats:sec>","DOI":"10.1093\/jac\/dkac046","type":"journal-article","created":{"date-parts":[[2022,2,8]],"date-time":"2022-02-08T20:11:30Z","timestamp":1644351090000},"page":"1296-1300","source":"Crossref","is-referenced-by-count":2,"title":["Spectrophotometric detection of azole-resistant <i>Aspergillus fumigatus<\/i> with the EUCAST broth microdilution method: is it time for automated MIC reading of EUCAST antifungal susceptibility testing of <i>Aspergillus<\/i> species?"],"prefix":"10.1093","volume":"77","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8869-1509","authenticated-orcid":false,"given":"Joseph","family":"Meletiadis","sequence":"first","affiliation":[{"name":"Clinical Microbiology Laboratory, Attikon University Hospital , Athens , Greece"},{"name":"Department of Medical Microbiology and Infectious Diseases, Erasmus Medical Center , Rotterdam , The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ioanna","family":"Efstathiou","sequence":"additional","affiliation":[{"name":"Clinical Microbiology Laboratory, Attikon University Hospital , Athens , Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hein A L","family":"van der Lee","sequence":"additional","affiliation":[{"name":"Department of Medical Microbiology, Radboud University Medical Centre, and Center of Expertise in Mycology Radboudumc\/CWZ , Nijmegen , The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Karen M T","family":"Astvad","sequence":"additional","affiliation":[{"name":"Unit of Mycology, Statens Serum Institut , Copenhagen , Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8600-9860","authenticated-orcid":false,"given":"Paul E","family":"Verweij","sequence":"additional","affiliation":[{"name":"Department of Medical Microbiology, Radboud University Medical Centre, and Center of Expertise in Mycology Radboudumc\/CWZ , Nijmegen , The Netherlands"},{"name":"Centre for Infectious Diseases Research, Diagnostics and Laboratory Surveillance, National Institute for Public Health and the Environment (RIVM) , Bilthoven , The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4747-0144","authenticated-orcid":false,"given":"Maiken Cavling","family":"Arendrup","sequence":"additional","affiliation":[{"name":"Unit of Mycology, Statens Serum Institut , Copenhagen , Denmark"},{"name":"Department of Clinical Microbiology, Rigshospitalet , Copenhagen , Denmark"},{"name":"Department of Clinical Medicine, University of Copenhagen , Copenhagen , Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2022,2,23]]},"reference":[{"key":"2023011417264633900_dkac046-B1","doi-asserted-by":"crossref","first-page":"1463","DOI":"10.1093\/cid\/ciy859","article-title":"Voriconazole resistance and mortality in invasive aspergillosis: a multicenter retrospective cohort study","volume":"68","author":"Lestrade","year":"2019","journal-title":"Clin Infect Dis"},{"article-title":"EUCAST DEFINITIVE DOCUMENT E.DEF 9.3.2. Method for the Determination of Broth Dilution Minimum Inhibitory Concentrations of Antifungal Agents for Conidia Forming Moulds","year":"2020","author":"Arendrup","key":"2023011417264633900_dkac046-B2"},{"volume-title":"Reference Method for Broth Dilution Antifungal Susceptibility Testing of Filamentous Fungi\u2014Second Edition: M38","year":"2008","author":"CLSI","key":"2023011417264633900_dkac046-B3"},{"key":"2023011417264633900_dkac046-B4","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1016\/j.cmi.2016.10.017","article-title":"Spectrophotometric reading of EUCAST antifungal susceptibility testing of Aspergillus fumigatus","volume":"23","author":"Meletiadis","year":"2017","journal-title":"Clin Microbiol Infect"},{"key":"2023011417264633900_dkac046-B5","doi-asserted-by":"crossref","first-page":"2431","DOI":"10.1128\/JCM.39.7.2431-2438.2001","article-title":"Identification of two different 14-\u03b1 sterol demethylase-related genes (cyp51A and cyp51B) in Aspergillus fumigatus and other Aspergillus species","volume":"39","author":"Mellado","year":"2001","journal-title":"J Clin Microbiol"},{"key":"2023011417264633900_dkac046-B6","doi-asserted-by":"crossref","first-page":"1464","DOI":"10.1016\/j.cmi.2020.06.007","article-title":"How to interpret MICs of antifungal compounds according to the revised clinical breakpoints v. 10.0 European Committee on Antimicrobial Susceptibility Testing (EUCAST)","volume":"26","author":"Arendrup","year":"2020","journal-title":"Clin Microbiol Infect"},{"key":"2023011417264633900_dkac046-B7","doi-asserted-by":"crossref","first-page":"478","DOI":"10.1128\/JCM.39.2.478-484.2001","article-title":"Analysis of growth characteristics of filamentous fungi in different nutrient media","volume":"39","author":"Meletiadis","year":"2001","journal-title":"J Clin Microbiol"},{"key":"2023011417264633900_dkac046-B8","doi-asserted-by":"crossref","first-page":"2053","DOI":"10.1128\/AAC.00873-06","article-title":"Concentration-dependent synergy and antagonism within a triple antifungal drug combination against Aspergillus species: analysis by a new response surface model","volume":"51","author":"Meletiadis","year":"2007","journal-title":"Antimicrob Agents Chemother"},{"key":"2023011417264633900_dkac046-B9","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1128\/AAC.47.1.106-117.2003","article-title":"In vitro drug interaction modeling of combinations of azoles with terbinafine against clinical Scedosporium prolificans isolates","volume":"47","author":"Meletiadis","year":"2003","journal-title":"Antimicrob Agents Chemother"},{"key":"2023011417264633900_dkac046-B10","doi-asserted-by":"crossref","first-page":"4256","DOI":"10.1128\/JCM.39.12.4256-4263.2001","article-title":"Comparison of spectrophotometric and visual readings of NCCLS method and evaluation of a colorimetric method based on reduction of a soluble tetrazolium salt, 2,3-bis {2-methoxy-4-nitro-5-[(sulfenylamino) carbonyl]-2H-tetrazolium-hydroxide}, for antifungal susceptibility testing of Aspergillus species","volume":"39","author":"Meletiadis","year":"2001","journal-title":"J Clin Microbiol"},{"key":"2023011417264633900_dkac046-B11","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1093\/jac\/dkg020","article-title":"In vitro susceptibilities of zygomycetes to conventional and new antifungals","volume":"51","author":"Dannaoui","year":"2003","journal-title":"J Antimicrob Chemother"},{"key":"2023011417264633900_dkac046-B12","doi-asserted-by":"crossref","first-page":"myab072","DOI":"10.1093\/mmy\/myab072","article-title":"Spectrophotometric azole and amphotericin B MIC readings against Aspergillus fumigatus sensu lato using the EUCAST 9.3.2 methodology. 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