title: Screening inhibitors of P. berghei blood stages using bioluminescent reporter parasites.
abstract: We describe two improved assays for in vitro and in vivo screening of inhibitors and chemicals for antimalarial activity against blood stages of the rodent malaria parasite, Plasmodium berghei. These assays are based on the determination of bioluminescence in small blood samples that is produced by reporter parasites expressing luciferase. Luciferase production increases as the parasite develops in a red blood cell and as the numbers of parasites increase during an infection. In the first assay, in vitro drug luminescence (ITDL) assay, the in vitro development of ring-stage parasites into mature schizonts in the presence and absence of candidate inhibitor(s) is quantified by measuring luciferase activity after the parasites have been allowed to mature into schizonts in culture. In the second assay, the in vivo drug luminescence (IVDL) assay, in vivo parasite growth (using a standard 4-day suppressive drug test) is quantified by measuring the luciferase activity of circulating parasites in samples of tail blood of drug-treated mice.
authors.affiliation: Center of Infectious Diseases, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Jing-Wen
authors.initials: JW
authors.lastname: Lin
authors.forename: Mohammed
authors.initials: M
authors.lastname: Sajid
authors.forename: Jai
authors.initials: J
authors.lastname: Ramesar
authors.forename: Shahid M
authors.initials: SM
authors.lastname: Khan
authors.forename: Chris J
authors.initials: CJ
authors.lastname: Janse
authors.forename: Blandine
authors.initials: B
authors.lastname: Franke-Fayard
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D000962:Antimalarials / Q000008:administration & dosage / Q000494:pharmacology*; D018929:Cell Culture Techniques; D004912:Erythrocytes / Q000187:drug effects / Q000469:parasitology*; D017930:Genes, Reporter; D008156:Luciferases / Q000235:genetics; D008163:Luminescent Measurements / Q000379:methods*; D008288:Malaria / Q000188:drug therapy / Q000469:parasitology; D051379:Mice; D021261:Parasitic Sensitivity Tests / Q000379:methods; D010962:Plasmodium berghei / Q000187:drug effects* / Q000254:growth & development
chemical_list: D000962:Antimalarials; D008156:Luciferases