Trypanosoma cruzi, Tulahuen CL2, b galactosidase strain (nifurtimox-sensitive) is used. All cultures and assays are conducted at 37°C under an atmosphere of 5% C02. Assays are performed in sterile 96-well microtiter plates, each well containing 10 µl of the watery compound dilutions together with 190 µl of MRC-5 cell/parasite inoculum (2.104 cells/ml + 2.105 parasites/ml). Parasite growth is compared to untreated-infected controls (100% growth) and non-infected controls (0% growth) after 7 days incubation. Parasite burdens are assessed after adding the substrate CPRG (chlorophenolred ß-D-galactopyranoside). The change in color is measured spectrophotometrically at 540 nm after 4 hours incubation at 37 °C. The results are expressed as % reduction in parasite burdens compared to control wells and an IC50 is calculated. Nifurtimox is included as reference drug.
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