A Dual-Readout High-Throughput Screening Assay for Small Molecules Active Against Aspergillus Fumigatus.

Human fungal infections caused by molds have been on the rise in recent years. These infections have high mortality rates compared to other fungal infections, and yet treatment options are limited due to resistance to clinical antifungals and lack of broad-spectrum activity against molds. Technical challenges associated with molds have limited large-throughput screening efforts for anti-mold compounds: therefore, we adapted an assay for use with A. fumigatus to help fill the gap in robust screening platforms for these organisms. This assay measures the release of the cytosolic enzyme adenylate kinase (AK) as a measure of fungal cell lysis and can also detect inhibition of germination as a reduction in the secretion of AK during vegetative growth. The ability to detect both lysis and inhibition of germination facilitates the identification of a wide range of compounds with different mechanisms of action, creating a strong screening platform for the identification of novel, anti-mold compounds.