“…Susceptibility (S), dose-dependent susceptibility (SDD) or resistance (R) were defined according to the CLSI breakpoints, which are for anidulafungin and caspofungin (in µg/mL): C. albicans S ≤ 0.25, R ≥ 1, C. glabrata S ≤ 0.12, R ≥ 0.5, for micafungin: C. albicans S ≤ 0.25, R ≥ 1, C. glabrata S ≤ 0.06, R ≥ 0.25, and for fluconazole: C. albicans S ≤ 2, R ≥ 8, C. glabrata SDD ≤ 32, R ≥ 64. Isolates for which MICs were confirmed to be above the susceptibility clinical breakpoints (i.e., classified as non-susceptible according to the CLSI breakpoints) for micafungin and/or anidulafungin were selected for genotypic characterization of FKS genes (of note, isolated elevated caspofungin MIC was not considered as a reliable predictor of FKS mutations on the basis of previous publications [10,12]). PCR and sequencing of the hotspot regions (HS) of the FKS genes (HS1 and HS2 of FKS1 for C. albicans and C. glabrata and HS1 and HS2 of FKS2 for C. glabrata) were performed using the primers previously described [10].…”