We tested 59 common and 27 uncommon Aspergillus species isolates for susceptibility to the mold-active azole antifungal agents itraconazole, voriconazole, and posaconazole using the Sensititre method. The overall essential agreement with the CLSI reference method was 96.5% for itraconazole and posaconazole and was 100% for voriconazole. By the Sensititre method as well as the CLSI reference method, all of 10 A. fumigatus isolates with a cyp51 mutant genotype were classified as being non-wild-type isolates (MIC Ͼ epidemiological cutoff value [ECV]) with respect to triazole susceptibility.KEYWORDS Aspergillus, Sensititre, antifungal susceptibility testing I n contrast to other but emerging molds (1), Aspergillus species, particularly Aspergillus fumigatus, remain the most common causes of invasive fungal diseases in both North America and Europe (2, 3). Because of the availability of (tri)azole antifungal agents, survival of immunocompromised patients with invasive aspergillosis has improved dramatically and could be further improved by optimizing antifungal treatments (4). A key component of this optimization should be the regular in vitro antifungal susceptibility testing of the patients' A. fumigatus isolates to detect azole resistance (5). Unfortunately, in most clinical microbiology laboratories, antifungal susceptibility testing of aspergilli (and other molds) is not routinely performed (6), thus underestimating the true prevalence of fungal resistance (4).The azole antifungal agents for clinical use include itraconazole, voriconazole, posaconazole, and, most recently, isavuconazole (7). Despite their role-unlike voriconazole, itraconazole and posaconazole are not approved as first-line agents-in treatment of invasive aspergillosis (4), the Clinical and Laboratory Standards Institute (CLSI) did not set clinical breakpoints (CBPs) for common Aspergillus species and mold-active triazoles, e.g., itraconazole and posaconazole (8), in contrast to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) (9). However, CLSI-based epidemiological cutoff values (ECVs) were established-instead of CBPs-for Aspergillus species (A. fumigatus, A. flavus, A. terreus, A. niger, A. nidulans, and A. versicolor) and for triazoles to aid in the early identification of clinical isolates with acquired resistance mechanisms (10, 11). Isolates of these six Aspergillus species for which triazole MICs exceed the ECV are considered to be non-wild type (non-WT) and may harbor mutations in the cyp51a gene-the best-known mechanism of triazole resistance in the A. fumigatus species-or other mutations (12). Interestingly, whereas the significance of ECVs in clinical practice needs to be understood, the ECVs defined to date-albeit mainly for Candida