Determination of antifungal drug susceptibilities of Aspergillus species by a fluorenscence-based microplate assay

S. Arunmozhi Balajee, Alexander Imhof, Jennifer L. Gribskov, Kieren A. Marr

Research output: Contribution to journalArticlepeer-review

Abstract

Objectives: We have investigated the use of a viability dye, chloromethylfluorescein di-acetate (CMFDA), for antifungal susceptibility testing in a fluorescence microplate (FM) assay format. Methods: For this FM assay, conidia were incubated in increasing concentrations of antifungal drug for 16h and stained with CMFDA. Fluorescence, measured as mean fluorescence units (MFU) in a fluorescence microplate reader, was graphed relative to that of a drug-free control, and the MIC was defined as the lowest concentration of the drug that resulted in complete reduction (100%) in MFU for amphotericin B, or 90% reduction in MFU for itraconazole and voriconazole. Susceptibilities of 10 clinical isolates of Aspergillus fumigatus, Aspergillus terreus and Aspergillus niger to amphotericin B, itraconazole and voriconazole were tested in a blinded fashion using the FM and the NCCLS methods. Results and conclusions: Reproducibility of the FM assay was excellent, and results correlated with those of the NCCLS microdilution method. The FM assay appears to be a rapid, objective method for testing fungal susceptibilities to itraconazole, voriconazole and amphotericin B.

Original languageEnglish (US)
Pages (from-to)102-105
Number of pages4
JournalJournal of Antimicrobial Chemotherapy
Volume55
Issue number1
DOIs
StatePublished - Jan 2005
Externally publishedYes

Keywords

  • Aspergillus spp.
  • Fluorescent dyes
  • MICS
  • Susceptibility testing

ASJC Scopus subject areas

  • Pharmacology
  • Microbiology (medical)
  • Infectious Diseases
  • Pharmacology (medical)

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