A Fluorescence-Based High-Throughput Screening Assay to Identify Growth Inhibitors of the Pathogenic Fungus Aspergillus fumigatus

Methods Mol Biol. 2016:1439:171-9. doi: 10.1007/978-1-4939-3673-1_11.

Abstract

Due to the advancements in modern medicine that have resulted in an increased number of immunocompromised individuals, the incidences and the associated mortality of invasive aspergillosis have continued to rise over the past three decades despite appropriate treatment. As a result, invasive aspergillosis has emerged as a leading cause of infection-related mortality in immunocompromised individuals. Utilizing the resazurin to resorufin conversion fluorescence readout to monitor cell viability, herein, we outline a high-throughput screening method amenable to profiling a large pharmaceutical library against the clinically relevant but less frequently screened fungal pathogen Aspergillus fumigatus. This enables the user to conduct high-throughput screening using a disease-relevant fungal growth assay and identify novel antifungal chemotypes as drug leads.

Keywords: Antifungal; Aspergillus fumigatus; Fungus; High-throughput screening (HTS); Resazurin.

MeSH terms

  • Antifungal Agents / pharmacology*
  • Aspergillosis / drug therapy*
  • Aspergillosis / microbiology
  • Aspergillus fumigatus / drug effects*
  • Aspergillus fumigatus / growth & development
  • Fluorescent Dyes / analysis
  • Fluorescent Dyes / metabolism
  • Growth Inhibitors / pharmacology*
  • High-Throughput Screening Assays / methods*
  • Humans
  • Microbial Sensitivity Tests / methods*
  • Oxazines / analysis
  • Oxazines / metabolism
  • Xanthenes / analysis
  • Xanthenes / metabolism

Substances

  • Antifungal Agents
  • Fluorescent Dyes
  • Growth Inhibitors
  • Oxazines
  • Xanthenes
  • resazurin