Evaluation of the Microdilution, Etest and Disk Diffusion Methods for Antifungal Susceptibility Testing of Clinical Strains of Trichosporon Spp.

Abstract

Trichosporon spp are well recognized as pathogens capable of causing invasive disease. Despite the increasing frequency and severity of trichosporonosis, data on the antifungal susceptibility of Trichosporon spp. are limited and recommendations for in vitro testing of this fungus are not included in the guidelines of the National Committee for Clinical Laboratory Standards. The purpose of this study was to determine the in vitro susceptibility of clinical Trichosporon isolates to systemic antifungals. We evaluated the in vitro activity of amphotericin B, fluconazole, itraconazole and voriconazole against 27 clinical isolates of Trichosporon spp. (14 T. mucoides and 13 T. asahii) using NCCLS M27-A2 reference microdilution, Etest and disk diffusion methods. In the microdilution and Etest methods Trichosporon spp. demonstrated relatively high minimum inhibitory concentrations (MICs) for fluconazole (MIC₉₀ 4 and 6 μg/ml, respectively) and relatively low MICs for voriconazole (MIC₉₀ 0.125 and 0.125 μg/ml, respectively). MICs for amphotericin B determined on antibiotic medium 3 were lower (MIC₉₀ 0.06 μg/ml) than those on RPMI (MIC₉₀ 1 μg/ml). Observed agreements were 81-100% according to these drugs. Disk diffusion zone diameters correlate inversely with MICs from dilution tests except for amphotericin B. Validation of the clinical significance of these observations demands determination of MIC breakpoints for Trichosporon and in vitro- in vivo correlation studies.

Keywords:

• Trichosporon spp
• antifungal susceptibility
• microdilution
• Etest
• disk diffusion
• amphotericin B
• fluconazole
• itraconazole
• voriconazole