Document Type
Article
Publication Date
3-17-2026
Original Citation
Flynn C,
Colón-Ríos R,
Harmez A,
Flynn W,
Kazmierczak B.
New Viability Assays for Acanthamoeba castellanii Trophozoites and Cysts Allow Rapid Assessment of Drug Activity. J Infect Dis. 2026;233(3):e766-e76.
Keywords
JGM, Acanthamoeba castellanii, Trophozoites, High-Throughput Screening Assays, Parasitic Sensitivity Tests, Drug Evaluation, Preclinical
JAX Source
J Infect Dis. 2026;233(3):e766-e76.
ISSN
1537-6613
PMID
41297556
DOI
https://doi.org/10.1093/infdis/jiaf599
Abstract
Pathogenic free-living amoebae such as Acanthamoeba castellanii are present in soil and water worldwide. A. castellanii causes systemic infections with very high mortality rates, yet drugs specifically targeting this pathogen are not available. Methods to reliably generate and assay cysts, which drive infection recurrence and drug resistance, are unavailable in a high-throughput format suitable for drug screening and testing. In this study, we developed a robust and reproducible protocol for encysting A. castellanii as well as a high-throughput, quantitative cyst viability assay using fluorescent live/dead staining coupled with microscopy and automated image analysis. These methods, coupled with optimized techniques for measuring trophozoite viability and pseudocyst formation, were used to screen the activity of 16 clinically relevant drugs and disinfectants. Four agents, including caspofungin, were active against both trophozoites and cysts.
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 4.0 International License.