Simple and sensitive antimalarial drug screening in vitro and in vivo using transgenic luciferase expressing Plasmodium berghei parasites.

摘要: We report two improved assays for in vitro and vivo screening of chemicals with potential anti-malarial activity against the blood stages rodent malaria parasite Plasmodiumberghei. These are based on determination luciferase (luminescence) small samples containing transgenic stage parasites that express under control a promoter is either schizont-specific (ama-1) or constitutive (eef1alphaa). Assay 1, drug luminescence (ITDL) assay, measured success schizont maturation presence candidate drugs quantifying mature schizonts only (ama-1 promoter). The ITDL assay generated drug-inhibition curves EC(50) values comparable to those obtained standard drug-susceptibility assays. second drug-luminescence (IVDL) growth 4-day suppressive test, monitored by measuring circulating (eef1alphaa IVDL generates growth-curves identical manual counting Giemsa-stained smears. reading rapid, requires minimal number handling steps no experience morphology fluorescence-activated cell sorters, produces radioactive waste test-plates can be stored prolonged periods before processing. Both tests suitable use larger-scale drugs. methodology validation, which includes testing models malaria, incorporation such