Luciferase-Based Growth Assay to Quantify Intracellular Parasite Reproduction
Published: April 30, 2023
Abstract
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)
In this video, we demonstrate the luciferase assay to quantify the growth of the intracellular pathogen Toxoplasma gondii. Utilizing a strain of Toxoplasma that expresses luciferase enzyme after infecting the host cells, the luciferase activity is detected in the infected cell culture to determine the successful reproduction of 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. Luciferase-based Toxoplasma growth assay Seed human foreskin fibroblasts (HFFs) 1 week before parasite inoculation to ensure that host cells are fully confluent. Perform a mock assay in a transparent plate to ensure that parasites remain intracellular throughout the evaluation period. NOTE: Here, the ass…
Divulgaciones
The authors have nothing to disclose.
Materials
| Biotek Synergy H1 Hybrid Multi-Mode Microplate Reader | BioTek Instuments | ||
| Coelenterazine h | Prolume | 301-10 hCTZ | |
| Software | |||
| GraphPad Prism software (8th version) |
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Luciferase-Based Growth Assay to Quantify Intracellular Parasite Reproduction. J. Vis. Exp. (Pending Publication), e21275, doi: (2023).