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We describe an in vitro microtest which is quantitative, rapid, and readily applicable to parasites isolated from all major forms of human leishmaniasis. It is based on drug-mediated inhibition of promastigote catabolism of a battery of simple 14C-substrates to 14CO2. Each test requires < 1µCi. The test is conducted in a serum-free, chemically defined medium containing 120 µg protein/ml, minimizing the possible interference of drug: serum protein interaction. Prior adaptation is not necessary to cultivate “difficult-to-grow” species. Leishmania sensitivity to pentavalent antimonials is detectable at µg levels below concentrations achievable in patients' sera.