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Evaluation of bioluminescence-based assays of anti-malarial drug activity
- Source :
- Malaria Journal, Vol 12, Iss 1, p 58 (2013), Malaria Journal
- Publication Year :
- 2013
- Publisher :
- BMC, 2013.
-
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. Conclusion 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.
- Subjects :
- Sybr green I
lcsh:Arctic medicine. Tropical medicine
Cell Survival
lcsh:RC955-962
Transgene
Plasmodium falciparum
lcsh:Infectious and parasitic diseases
QH301
03 medical and health sciences
chemistry.chemical_compound
Antimalarials
Inhibitory Concentration 50
Parasitic Sensitivity Tests
Antimalarial drug
Genes, Reporter
parasitic diseases
Bioluminescence
Humans
Luciferase
lcsh:RC109-216
Luciferases
030304 developmental biology
0303 health sciences
biology
Staining and Labeling
030306 microbiology
Research
Lethal dose
biology.organism_classification
Relative rate of kill
Molecular biology
Fluorescence
3. Good health
Malaria
Infectious Diseases
Parasitology
chemistry
Luminescent Measurements
SYBR Green I
Subjects
Details
- Language :
- English
- ISSN :
- 14752875
- Volume :
- 12
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Malaria Journal
- Accession number :
- edsair.doi.dedup.....52e628eb09a5a3e8803d7f50219de5bb