Comparison of a new commercial colorimetric microdilution method with a standard method for in-vitro susceptibility testing of Candida spp. and Cryptococcus neoformans

Abstract

The Sensititre Yeast One method (AccuMed International Ltd, East Grinstead, UK) is a microplate-based procedure that incorporates an oxidation-reduction indicator, Alamar Blue, for the in-vitro testing of five antifungal agents (amphotericin B, fluconazole, itraconazole, ketoconazole and flucytosine). We compared this colorimetric method with a standard broth microdilution test, performed according to US National Committee for Clinical Laboratory Standards document M27-A guidelines, for determining the in-vitro susceptibilities of 180 isolates of Candida spp. (50 Candida albicans, 50 Candida glabrata, ten Candida kefyr, 20 Candida krusei, ten Candida lusitaniae, 20 Candida parapsilosis and 20 Candida tropicalis) and 20 isolates of Cryptococcus neoformans. The overall agreement between the results of the two methods (colorimetric MICs within +/- two log2 dilutions of standard MICs) were 99% for amphotericin B, 96.5% for flucytosine, 93% for itraconazole, 91.5% for fluconazole and 85.5% for ketoconazole. The overall levels of agreement between the two methods were > or = 94% for six of the eight species tested, the exceptions being C. neoformans and C. tropicalis where the overall agreement was 89% and 80% respectively. The poorest agreement between the results for individual agents was seen with C. tropicalis and the three azole agents (60-75% of colorimetric MICs within +/- two log2 dilutions of standard MICs), and C. neoformans for ketoconazole (50%). The Yeast One method appears to be a suitable alternative procedure for routine antifungal susceptibility testing of Candida spp. and C. neoformans.