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In?vitro susceptibility was determined by following the guidelines in the AFST-EUCAST definitive document?7.1 [8] or a modified EUCAST technique, using AM3 medium in place of RPMI for amphotericin?B and caspofungin. Microplates were prepared in batches and stored frozen at ?20��C for [http://www.selleckchem.com/products/mcc950-sodium-salt.html MCC950 in vivo] of the determination by the EUCAST method at the NRCMA. All laboratories used [http://en.wikipedia.org/wiki/PRDX5 PRDX5] RPMI agar with 2% glucose, and MICs were determined when growth was clearly seen after 24�C48?h, as recommended by the manufacturer (AB Biodisk, Solna, Sweden). Some hospitals did not test all of the available Etest antifungal strips on each isolate. However, missing values were not obtained retrospectively. Low off-scale values were left unchanged, and high off-scale values were converted to the next higher concentration. For calculations, the Etest MICs were raised to the next corresponding EUCAST concentration. Agreement was defined as differences of no more than two dilutions between results obtained by the two techniques. Categorical agreement was defined as the percentage of isolates classified into the same category with both methods, based on breakpoints previously published for all antifungals except caspofungin [11,19,20]. Tentative interpretative breakpoints were those recommended by EUCAST for fluconazole [19] (susceptible ��2?mg/L, intermediate 4?mg/L, and resistant ��8?mg/L) and for voriconazole [20] (susceptible [http://www.selleckchem.com/products/midostaurin-pkc412.html click here] ��0.125?mg/L, and resistant ��0.25?mg/L), and those used previously [11] for amphotericin?B (susceptible ��0.25?mg/L, intermediate 0.50�C1.0?mg/L, and resistant ��2?mg/L), flucytosine (susceptible ��4?mg/L, intermediate 8�C16?mg/L, and resistant ��32?mg/L), and itraconazole (susceptible ��0.12?mg/L, intermediate 0.25�C0.50?mg/L, and resistant ��1?mg/L). For caspofungin, on the basis of MIC determinations for wild-type and fks1 mutant isolates [21], the following breakpoints were used: susceptible ��0.25?mg/L, and resistant ��0.5?mg/L [22]. Discrepancies were considered to be very major if an isolate classified as resistant by the reference method was categorized as susceptible by the commercial method. Discrepancies were considered to be major if an isolate classified as susceptible by the reference method was classified as resistant by the commercial technique. Errors were classified as minor when susceptible vs. intermediate, resistant vs.