title: Evaluation of bioluminescence-based assays of anti-malarial drug activity.
abstract: BACKGROUND
  Transgenic Plasmodium falciparum expressing luciferase offers an attractive bioluminescence-based assay platform for the investigation of the pharmacological properties of anti-malarial drugs. Here a side-by-side comparison of bioluminescence and fluorescence-based assays, utilizing a luciferase reporter cassette that confers a strong temporal pattern of luciferase expression during the S-phase of intraerythrocytic development, is reported.
  
  
  METHODS
  Key assay parameters for a range of commercially available luminogenic substrates are determined and compared to those measured using a Malaria Sybr Green I fluorescence assay. In addition, the short-term temporal effects of anti-malarial compounds are evaluated using both bioluminescent and fluorescent assay platforms.
  
  
  RESULTS
  The Z', % coefficient of variation and 50% inhibition concentrations are essentially the same for bioluminescent and fluorescent assays in transgenic parasites generated in both chloroquine-sensitive and -resistant genetic backgrounds. Bioluminescent assays, irrespective of the luminogenic agent employed, do, however, offer significantly enhanced signal-to-noise ratios. Moreover, the bioluminescent assay is more dynamic in terms of determining temporal effects immediately following drug perturbation.
  
  
  CONCLUSIONS
  This study suggests that opportunities for bioluminescence-based assays lie not in the measurement of 50% inhibition concentrations, where the cheaper fluorescence assay performs excellently and is not restricted by the need to genetically modify the parasite clone under investigation. Instead, assays that use the dynamic response of the luciferase reporter for semi-automated screening of additional pharmacological properties, such as relative rate-of-kill and lethal dose estimation, are a more attractive development opportunity.
authors: Institute for Science and Technology in Medicine, Keele University, Staffordshire, UK.
authors: Sandra
authors: S
authors: Hasenkamp
authors: Adam
authors: A
authors: Sidaway
authors: Oliver
authors: O
authors: Devine
authors: Richard
authors: R
authors: Roye
authors: Paul
authors: P
authors: Horrocks
mesh_terms: D000962:Antimalarials / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D017930:Genes, Reporter; D006801:Humans; D020128:Inhibitory Concentration 50; D008156:Luciferases / Q000032:analysis / Q000235:genetics; D008163:Luminescent Measurements / Q000379:methods*; D021261:Parasitic Sensitivity Tests / Q000379:methods*; D010963:Plasmodium falciparum / Q000187:drug effects*; D013194:Staining and Labeling / Q000379:methods
chemical_list: D000962:Antimalarials; D008156:Luciferases
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