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In vitro antifungal susceptibility testing of filamentous fungi with Sensititre Yeast OneTM

Sensititre® is a colorimetric microdilution method for in vitro antifungal susceptibility testing based on the M27‐A document (National Committee for Clinical Laboratory Standards) for yeasts. Difference between both methods is the presence of Alamar‐blue® and RPMI 1640 (glucose 2%) as culture medium. Antifungal susceptibility to amphotericin B, fluconazole, itraconazole, ketoconazole and flucytosine, 100 opportunistic filamentous fungi (Aspergillus spp., Fusarium spp., Scedosporium spp.) obtained from pathological samples was determined by the Sensititre method. Induction to conidium and sporangiospore formation at 35°C was used to get inoculum and plates were covered by 1 ml of saline and suspensions were made by gently probing by a sterile loop. Optical densities of the conidial suspensions were adjusted to 80–82% transmittance for Aspergillus spp. and 68–70% for the rest of strains tested. Final inoculum concentration size was 0.4 × 104–5 × 104 CFU ml−1. Readings were made at 72 h of incubation at 35°C; amphotericin B and itraconazole was active against Aspergillus fumigatus with CMI90 1 and 0.5 μg ml−1, respectively, opposite to Scedosporium prolificans and Scedosporium apiospermum. As it was expected, a CMI90 of 256 μg ml−1 for fluconazole and CMI90 for flucytosine amounting to 64 g ml−1 were obtained. Sensititre Yeast OneTM is a useful method and an alternative to reference methods to determine antifungal susceptibility of filamentous fungi for clinical laboratory routine. Correlation with microdilution results is studied. New triazole derivatives should be included as soon as their clinical use will be feasible.