Evaluation of the Etest Method Using Mueller-Hinton Agar with Glucose and Methylene Blue for Determining Amphotericin B MICs for 4,936 Clinical Isolates of Candida Species

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Abstract
The performance of the Etest using Mueller-Hinton agar supplemented with glucose (2%) and methylene blue (0.5 μg/ml) (MH-GMB) for amphotericin B susceptibility testing of 4,936 isolates of Candida spp. was assessed against that of Etest using RPMI agar with 2% glucose (RPG). MICs were determined by Etest in both media for all 4,936 isolates and were read after incubation for 48 h at 35°C. The Candida isolates included C. albicans ( n = 2,728), C. glabrata ( n = 722), C. parapsilosis ( n = 666), C. tropicalis ( n = 528), C. krusei ( n = 143), C. lusitaniae ( n = 54), C. guilliermondii ( n = 39), C. pelliculosa ( n = 17), C. kefyr ( n = 15), C. rugosa ( n = 11), C. dubliniensis ( n = 5), C. zeylanoides ( n = 4), C. lipolytica ( n = 3), and C. famata ( n = 1). The Etest results with MH-GMB correlated well with those with RPG. Overall agreement was 92.9%, and agreements for individual species were as follows: C. lusitaniae , 98.1%; C. albicans , 95.1%; C. glabrata , 94.3%; C. krusei , 91.6%; C. parapsilosis , 86.6%; and C. tropicalis , 86.4%. The Etest method using MH-GMB appears to be a useful method for determining amphotericin B susceptibilities of Candida species.

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