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Luciferase Assay to Study Efficacy of Antiparasitic Chemicals Against Parasites
Overview
The video demonstrates the luciferase assay used to determine the efficacy of an antiparasitic chemical inhibitor against the intracellular pathogen Toxoplasma gondii. The study involves luciferase-expressing parasites invading the host cells to replicate and survive. The chemical inhibitor binds with the cysteine proteases of the parasite and inhibits replication. The luciferase activity detected is proportional to the fold changes in parasite growth, determining the drug's efficacy against the parasite.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Evaluation of chemical compound inhibition efficacy against Toxoplasma growth
NOTE: Here, evaluation of the inhibition of LHVS in Toxoplasma growth is presented as an example. Eight different concentrations of LHVS are tested, and three technical replicates are performed for each of the t.......
Representative Results
Figure 1: Inhibition efficacy assessment of LHVS and pyrimethamine using the luciferase-based growth assay. Parasites were inoculated into a 96-well microplate for 4 h to allow for invasion of host cells. Non-invaded parasites were washed away, and the plate was filled with media containing different concentrations of LHVS or pyrimethamine and incubated for an additional 96 h before determination.......
This article has been published
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Source: Key, M., et al. Determination of Chemical Inhibitor Efficiency against Intracellular Toxoplasma Gondii Growth Using a Luciferase-Based Growth Assay. J. Vis. Exp. (2020).
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