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Dual-Luciferase-Based Fast and Sensitive Detection of Malaria Hypnozoites for the Discovery of Antirelapse Compounds
- Source :
- Analytical Chemistry
- Publication Year :
- 2020
-
Abstract
- Efforts to eradicate Plasmodium vivax malaria are hampered by the presence of hypnozoites, persisting stages in the liver that can reactivate after prolonged periods of time enabling further transmission and causing renewed disease. Large-scale drug screening is needed to identify compounds with antihypnozoite activity, but current platforms rely on time-consuming high-content fluorescence imaging as read-out, limiting assay throughput. We here report an ultrafast and sensitive dual-luciferase-based method to differentiate hypnozoites from liver stage schizonts using a transgenic P. cynomolgi parasite line that contains Nanoluc driven by the constitutive hsp70 promoter, as well as firefly luciferase driven by the schizont-specific lisp2 promoter. The transgenic parasite line showed similar fitness and drug sensitivity profiles of selected compounds to wild type. We demonstrate robust bioluminescence-based detection of hypnozoites in 96-well and 384-well plate formats, setting the stage for implementation in large scale drug screens.
- Subjects :
- Plasmodium
Transgene
010402 general chemistry
01 natural sciences
Article
Analytical Chemistry
Antimalarials
Parasitic Sensitivity Tests
parasitic diseases
Drug Discovery
medicine
Bioluminescence
Animals
Luciferase
Luciferases
Cells, Cultured
Liver stage
Chemistry
Drug screens
010401 analytical chemistry
Optical Imaging
Wild type
medicine.disease
Virology
Macaca mulatta
0104 chemical sciences
Malaria
Luminescent Measurements
Hepatocytes
Plasmodium vivax Malaria
Subjects
Details
- ISSN :
- 15206882
- Volume :
- 92
- Issue :
- 9
- Database :
- OpenAIRE
- Journal :
- Analytical chemistry
- Accession number :
- edsair.doi.dedup.....3f106f42c01a250f8850a814278cf8dc