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Citation
KARL, S., WONG, R.P., ST PIERRE, T.G. & DAVIS, T.M.E. (2009). A comparative study of a flow-cytometry-based assessment of in vitro Plasmodium falciparum drug sensitivity. Malaria Journal, 8 (294), 1-11.
Title
A comparative study of a flow-cytometry-based assessment of in vitro Plasmodium falciparum drug sensitivity
Author
Year
2009
Published in
Abstract
BackgroundRecently developed Sybr Green-based in vitro Plasmodium falciparum drug sensitivity assays provide an attractive alternative to current manual and automated methods. The present study evaluated flow cytometry measurement of DNA staining with Sybr Green in comparison with the P. falciparum lactate dehydrogenase assay, the tritiated hypoxanthine incorporation assay, a previously described Sybr Green based plate reader assay and light microscopy.MethodsAll assays were set up in standardized format in 96-well plates. The 50% inhibitory concentrations (IC50) of chloroquine, mefloquine and dihydroartemisinin against the laboratory adapted P. falciparum strains 3D7, E8B, W2mef and Dd2 were determined using each method.ResultsThe resolution achieved by flow cytometry allowed quantification of the increase in individual cell DNA content after an incubation period of only 24 h. Regression, and Bland and Altman analyses showed that the IC50 values determined using the flow cytometry assay after 24 h agreed well with those obtained using the hypoxanthine incorporation assay, the P. falciparum lactate dehydrogenase assay, the Sybr Green plate reader assay and light microscopy. However the values obtained with the flow cytometry assay after 48 h of incubation differed significantly from those obtained with the hypoxanthine incorporation assay, and the P. falciparum lactate dehydrogenase assay at low IC50 values, but agreed well with the Sybr Green plate reader assay and light microscopy.ConclusionsAlthough flow cytometric equipment is expensive, the necessary reagents are inexpensive, the procedure is simple and rapid, and the cell volume required is minimal. This should allow field studies using fingerprick sample volumes.
Identifier
http://dx.doi.org/10.1186/1475-2875-8-294
Identifier
KARL, S., WONG, R.P., ST PIERRE, T.G. & DAVIS, T.M.E. (2009). A comparative study of a flow-cytometry-based assessment of in vitro Plasmodium falciparum drug sensitivity. Malaria Journal, 8 (294), 1-11.
Identifier
1475-2875
Identifier
http://repository.uwa.edu.au:80/R/-?func=dbin-jump-full&object_id=27933&silo_library=GEN01
ISBN/ISSN
1475-2875
Subject
Department/School
Type
Journal article
DOI
Persistent URL
http://repository.uwa.edu.au:80/R/-?func=dbin-jump-full&object_id=27933&silo_library=GEN01
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Life and Physical Sciences > Physics
Medicine, Dentistry and Health Sciences > Medicine and Pharmacology