ABSTRACT The CLSI epidemiological cutoff values (ECVs) of antifungal agents are available for various Candida spp., Aspergillus spp., and the Mucorales. However, those categorical endpoints have not been established for Fusarium spp., mostly due to the difficulties associated with collecting sufficient CLSI MICs for clinical isolates identified according to the currently recommended molecular DNA-PCR-based identification methodologies. CLSI MIC distributions were established for 53 Fusarium dimerum species complex (SC), 10 F. fujikuroi , 82 F. proliferatum , 20 F. incarnatum-F. equiseti SC, 226 F. oxysporum SC, 608 F. solani SC, and 151 F. verticillioides isolates originating in 17 laboratories (in Argentina, Australia, Brazil, Canada, Europe, Mexico, and the United States). According to the CLSI guidelines for ECV setting, ECVs encompassing ≥97.5% of pooled statistically modeled MIC distributions were as follows: for amphotericin B, 4 μg/ml ( F. verticillioides ) and 8 μg/ml ( F. oxysporum SC and F. solani SC); for posaconazole, 2 μg/ml ( F. verticillioides ), 8 μg/ml ( F. oxysporum SC), and 32 μg/ml ( F. solani SC); for voriconazole, 4 μg/ml ( F. verticillioides ), 16 μg/ml ( F. oxysporum SC), and 32 μg/ml ( F. solani SC); and for itraconazole, 32 μg/ml ( F. oxysporum SC and F. solani SC). Insufficient data precluded ECV definition for the other species. Although these ECVs could aid in detecting non-wild-type isolates with reduced susceptibility to the agents evaluated, the relationship between molecular mechanisms of resistance (gene mutations) and MICs still needs to be investigated for Fusarium spp.