Format

Send to

Choose Destination
Anal Bioanal Chem. 2013 Aug;405(21):6823-9. doi: 10.1007/s00216-013-7134-4. Epub 2013 Jun 30.

A high-throughput screening assay for assessing the viability of Cryptococcus neoformans under nutrient starvation conditions.

Author information

1
National Center for Advancing Translational Sciences, National Institute of Health, 9800 Medical Center Drive, MSC: 3370, Bethesda, MD 20892-3370, USA.

Abstract

Cryptococcus neoformans causes an estimated 600,000 AIDS-related deaths annually that occur primarily in resource-limited countries. Fluconazole and amphotericin B are currently available for the treatment of cryptococcal-related infections. However, fluconazole has limited clinical efficacy and amphotericin B requires intravenous infusion and is associated with high renal toxicity. Therefore, there is an unmet need for a new orally administrable anti-cryptococcal drug. We have developed a high-throughput screening assay for the measurement of C. neoformans viability in 1,536-well plate format. The signal-to-basal ratio of the ATP content assay was 21.9 fold with a coefficient of variation and Z' factor of 7.1% and 0.76, respectively. A pilot screen of 1,280 known compounds against the wild-type C. neoformans (strain H99) led to the identification of four active compounds including niclosamide, malonoben, 6-bromoindirubin-3'-oxime, and 5-[(4-ethylphenyl)methylene]-2-thioxo-4-thiazolidinone. These compounds were further tested against nine clinical isolates of C. neoformans, and their fungicidal activities were confirmed. The results demonstrate that this miniaturized C. neoformans assay is advantageous for the high-throughput screening of large compound collections to identify lead compounds for new anti-cryptococcal drug development.

PMID:
23812880
PMCID:
PMC4539003
DOI:
10.1007/s00216-013-7134-4
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Springer Icon for PubMed Central
Loading ...
Support Center