Modification of the fluorescein diacetate assay for screening of antifungal agents against Candida albicans: Comparison with the NCCLS methods

Nynke Brouwer, James Kohen, Joanne Jamie, Subramanyam Vemulpad

Research output: Contribution to journalArticleResearchpeer-review

Abstract

A modified fluorescein diacetate (FDA) assay has been compared with standard NCCLS broth macrodilution and broth microdilution methods for the detection of antifungal activity. The FDA assay was performed in a medium containing bacteriological peptone, NaCl, yeast extract and glucose (0.2%, 0.1%, 0.1% and 1% w/v, respectively) and buffered with 10 mM BES buffer. The MICs of amphotericin B, fluconazole, miconazole and flucytosine (representing three major classes of antifungal agents) obtained by the three methods were compared. The results obtained with the FDA assays correlated well with the NCCLS macrodilution method for MICs of amphotericin B, miconazole and fluconazole, but not for flucytosine. However, the MIC values of flucytosine obtained with the FDA assay were well within the quality control range for the two reference strains recommended by the NCCLS. The FDA assay described is an attractive alternative to the NCCLS methods for screening for antifungal agents, with the added advantage of objectivity of fluorescence measurement.

LanguageEnglish
Pages234-241
Number of pages8
JournalJournal of Microbiological Methods
Volume66
Issue number2
DOIs
Publication statusPublished - Aug 2006

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Antifungal Agents
Candida albicans
Flucytosine
Miconazole
Fluconazole
Amphotericin B
Peptones
Quality Control
Buffers
Reference Values
Yeasts
Fluorescence
3',6'-diacetylfluorescein
Glucose

Cite this

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title = "Modification of the fluorescein diacetate assay for screening of antifungal agents against Candida albicans: Comparison with the NCCLS methods",
abstract = "A modified fluorescein diacetate (FDA) assay has been compared with standard NCCLS broth macrodilution and broth microdilution methods for the detection of antifungal activity. The FDA assay was performed in a medium containing bacteriological peptone, NaCl, yeast extract and glucose (0.2{\%}, 0.1{\%}, 0.1{\%} and 1{\%} w/v, respectively) and buffered with 10 mM BES buffer. The MICs of amphotericin B, fluconazole, miconazole and flucytosine (representing three major classes of antifungal agents) obtained by the three methods were compared. The results obtained with the FDA assays correlated well with the NCCLS macrodilution method for MICs of amphotericin B, miconazole and fluconazole, but not for flucytosine. However, the MIC values of flucytosine obtained with the FDA assay were well within the quality control range for the two reference strains recommended by the NCCLS. The FDA assay described is an attractive alternative to the NCCLS methods for screening for antifungal agents, with the added advantage of objectivity of fluorescence measurement.",
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Modification of the fluorescein diacetate assay for screening of antifungal agents against Candida albicans : Comparison with the NCCLS methods. / Brouwer, Nynke; Kohen, James; Jamie, Joanne; Vemulpad, Subramanyam.

In: Journal of Microbiological Methods, Vol. 66, No. 2, 08.2006, p. 234-241.

Research output: Contribution to journalArticleResearchpeer-review

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