title: Interlaboratory evaluation of Etest method for testing antifungal susceptibilities of pathogenic yeasts to five antifungal agents by using Casitone agar and solidified RPMI 1640 medium with 2% glucose.
abstract: An interlaboratory evaluation (two centers) of the Etest method was conducted for testing the antifungal susceptibilities of yeasts. The MICs of amphotericin B, fluconazole, flucytosine, itraconazole, and ketoconazole were determined for 83 isolates of Candida spp., Cryptococcus neoformans, and Torulopsis glabrata. Two buffered (phosphate buffer) culture media were evaluated: solidified RPMI 1640 medium with 2% glucose and Casitone agar. MIC endpoints were determined after both 24 and 48 h of incubation at 35 degrees C. Analysis of 3,420 MICs demonstrated higher interlaboratory agreement (percentage of MIC pairs within a 2-dilution range) with Casitone medium than with RPMI 1640 medium when testing amphotericin B (84 to 90% versus 1 to 4%), itraconazole (87% versus 63 to 74%), and ketoconazole (94 to 96% versus 88 to 90%). In contrast, better interlaboratory reproducibility was determined between fluconazole MIC pairs when RPMI 1640 medium rather than Casitone medium was used (96 to 98% versus 77 to 90%). Comparison of the flucytosine MICs obtained with RPMI 1640 medium revealed greater than 80% reproducibility. The study suggests the potential value of the Etest as a convenient alternative method for testing the susceptibilities of yeasts. It also indicates the need for further optimization of medium formulations and MIC endpoint criteria to improve interlaboratory agreement.
authors.affiliation: Medical College of Virginia/Virginia Commonwealth University, Richmond 23298-0049, USA.
authors.forename: A
authors.initials: A
authors.lastname: Espinel-Ingroff
authors.forename: M
authors.initials: M
authors.lastname: Pfaller
authors.forename: M E
authors.initials: ME
authors.lastname: Erwin
authors.forename: R N
authors.initials: RN
authors.lastname: Jones
mesh_terms: D000362:Agar; D000935:Antifungal Agents / Q000494:pharmacology*; D002175:Candida / Q000187:drug effects; D002364:Caseins; D003455:Cryptococcus neoformans / Q000187:drug effects; D003470:Culture Media; D005069:Evaluation Studies as Topic; D005947:Glucose; D006801:Humans; D007753:Laboratories; D008826:Microbial Sensitivity Tests / Q000379:methods* / Q000592:standards / Q000706:statistics & numerical data; D011492:Protein Hydrolysates; D012015:Reference Standards; D015203:Reproducibility of Results; D015003:Yeasts / Q000187:drug effects* / Q000302:isolation & purification / Q000472:pathogenicity
chemical_list: D000935:Antifungal Agents; D002364:Caseins; D003470:Culture Media; D011492:Protein Hydrolysates; C026375:trypticase-soy broth; D000362:Agar; D005947:Glucose