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Olorofim is a novel antifungal agent within vitro activity againstAspergillus and some other molds. Here, we addressed technical aspects for EUCAST olorofim testing and generated contemporary MIC data. EUCAST E.Def 9.3.1 testing was performed comparing two plate preparation methods (serial dilution in medium [serial plates] versus predilution in DMSO [ISO plates]), two lots of olorofim, visual (visual-MIC) versus spectrophotometer (spec-MIC) reading, and four polystyrene plates using 34 to 53Aspergillus isolates from five genera. Subsequently, olorofim MICs were compared to itraconazole, voriconazole, posaconazole, and amphotericin B MICs for 298 clinical mold isolates (2016 to 2017). Wild-type upper limits (WT-UL) were determined following EUCAST principles for epidemiologic cutoff value (ECOFF) setting. Olorofim median MICs comparing serial plates and ISO plates were identical (25/36 [69%]) or one dilution apart (11/36 [31%]). Interperson agreement for visual-MICs was 92% to 94%/100% for ≤1/≤2 dilutions, respectively. The visual-MIC values across tested microtiter plates and olorofim lots revealed only discrete differences (≤1 dilution lower for treated plates). No single spec-MIC criterion was applicable to all species. Olorofim MICs were low against 275Aspergillus species isolates (modal MIC, 0.06 mg/liter; MIC range, &lt; 0.004 to 0.25 mg/liter) and three dermatophytes (MICs 0.03 to 0.06 mg/liter). MICs againstFusarium were diverse, with full inhibition ofF. proliferatum (MIC, 0.016), 50% growth inhibition ofFusarium solani at 1 to 2 mg/liter, and no inhibition ofF. dimerum . Olorofim displayed potentin vitro activity against most mold isolates and was associated with limited variation in EUCAST susceptibility testing.

EUCAST Determination of Olorofim (F901318) Susceptibility of Mold Species, Method Validation, and MICs