Toxoplasma gondii: A simple high-throughput assay for drug screening in vitro

Chun Mei Jin, Kusuma Kaewintajuk, Jing Hua Jiang, Woo Jin Jeong, Masaki Kamata, Hye Sook KIM, Yusuke Wataya, Hyun Park

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Toxoplasma gondii is the etiologic agent of toxoplasmosis. Although the combination of sulfadiazine and pyrimethamine is used as therapy for this disease, these drugs can have serious side effects and its use is limited in pregnancy. Therefore there is a need for new anti-T. gondii drugs in the clinic. Some systems for T. gondii drug screening have been described, but these have limitations and can be difficult. In order to solve these problems, we established a system to screen drugs in vitro that involved using cell viability methods to calculate drug selectivities, which are Trypan blue, [3-(4,5-dimethylthiazol-zyl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazoliuzolium, inner salt] (MTS) method and lactate dehydrogenase (LDH) assay. These assays were simple to establish and perform. The IC50 values calculated from the morphological assay were not significantly different from the EC50 values calculated using the other three methods. In particular, the results of the morphological assay showed a distinct association with the MTS assay (R = 0.9841). These assays could be used for a wide range of applications in the screening of new drugs and may provide an alternative to the techniques currently used to screen for candidate anti-T. gondii compounds in vitro. In this study, we also tested many compounds and identified some that had a good anti-T. gondii effect in vitro based on the MTS assay. This simple and fast system allowed us to determine which compounds to investigate further using in vivo experiments.

Original languageEnglish
Pages (from-to)132-136
Number of pages5
JournalExperimental Parasitology
Volume121
Issue number2
DOIs
Publication statusPublished - Feb 1 2009

Keywords

  • LDH assay
  • MTS method
  • Toxoplasma gondii drug screening system
  • Trypan blue dye

ASJC Scopus subject areas

  • Parasitology
  • Immunology
  • Infectious Diseases

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  • Cite this

    Jin, C. M., Kaewintajuk, K., Jiang, J. H., Jeong, W. J., Kamata, M., KIM, H. S., Wataya, Y., & Park, H. (2009). Toxoplasma gondii: A simple high-throughput assay for drug screening in vitro. Experimental Parasitology, 121(2), 132-136. https://doi.org/10.1016/j.exppara.2008.10.006