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J Biomol Screen. 2014 Feb;19(2):270-277. doi: 10.1177/1087057113496847. Epub 2013 Jul 29.

A high-throughput screening assay for fungicidal compounds against Cryptococcus neoformans.

Author information

Laboratory of Clinical Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA.
Department of Laboratory Medicine, Clinical Center, National Institutes of Health, Bethesda, MD USA.
Chemistry Department, Georgetown University, Washington D.C., USA.
Section of Infectious Diseases, Department of Medicine, University of Illinois at Chicago, IL, USA.
Contributed equally


Cryptococcus neoformans is a pathogenic fungus that causes meningitis worldwide, particularly in human immunodeficiency virus (HIV)-infected individuals. Although amphotericin B is the "gold standard" treatment for cryptococcal meningitis, the toxicity and inconvenience of intravenous injection emphasize a need for development of new anticryptocccal drugs. Recent data from humans and animal studies suggested that a nutrient-deprived host environment may exist in cryptococcal meningitis. Thus, a screening assay for identifying fungicidal compounds under nutrient-deprived conditions may provide an alternative strategy to develop new anticryptococcal drugs for this disease. A high-throughput fungicidal assay was developed using a profluorescent dye, alamarBlue, to detect residual metabolic activity of C. neoformans under nutrient-limiting conditions. Screening the Library of Pharmacologically Active Compounds (LOPAC) with this assay identified a potential chemical scaffold, 10058-F4, that exhibited fungicidal activity in the low micromolar range. These results thus demonstrate the feasibility of this alamarBlue-based assay for high-throughput screening of fungicidal compounds under nutrient-limiting conditions for new anticryptococcal drug development.


Cryptococcus neoformans; alamarBlue assay; fungicidal screen; high-throughput screen

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