Microtiter broth dilution method for yeast susceptibility testing with validation by clinical outcome.

Journal of Clinical Microbiology
M RadetskyJ K Todd

Abstract

There is no ideal laboratory procedure or culture medium in current use for susceptibility testing of pathogenic yeasts. Six candidate growth media (RPMI 1640 with L-glutamine, yeast nitrogen base, Casamino Acids medium, Mueller-Hinton broth, Sabouraud dextrose broth, and minimum essential medium-Eagle salts) were screened by spectrophotometric absorbance for nucleic acid and protein. From these, two media were selected: a chemically defined growth medium (RPMI 1640 with L-glutamine) and a chemically complex medium (Casamino Acids). MICs of four antifungal agents (5-fluorocytosine, miconazole, ketoconazole, and amphotericin B) for 84 clinical isolates of various Candida species were then determined with both media in agar dilution and microtiter broth dilution systems. The resultant MICs were correlated with clinical outcome for those isolates obtained from patients treated with single antifungal agents, and susceptibility cut points were calculated. Derived MIC cut points for susceptibility were validated in a murine model of systemic candidiasis. RPMI 1640 with L-glutamine was found to have the lowest absorbance values for both nucleic acid and protein, while Casamino Acids medium was highest in both categories. We found that...Continue Reading

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