by Yang G, Lee N, Ioset JR, No JH. Journal of Biomolecular Screening 2016, pii: 1087057116673796.
Summary: In this study, different Trypanosoma cruzi assay protocols were evaluated in order to understand the key parameters influencing drug susceptibility. Two assays were compared, one an image-based intracellular T. cruzi assay and the other utilizing a β-galactosidase-expressing T. cruzi strain. Initial comparisons of the assays with 31 reference compounds showed a very poor correlation. Nitroheterocyclics and CYP51 inhibitors were inactive in an image-based assay, but were highly active in a colorimetric assay. A high correlation was observed only after both compound incubation time and cell seeding were synchronized between assays. Further analysis showed that nitroheterocyclics and CYP51 inhibitors exhibit relatively large deviations in activity between experimental protocols routinely used for in vitro intracellular T. cruzi assays, suggesting that the factors mentioned are critical in their design.