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Title: CyQUANT cell proliferation assay as a fluorescence-based method for in vitro screening of antimalarial activity
Authors: Nongluk Sriwilaijaroen
Jane Xu Kelly
Michael Riscoe
Prapon Wilairat
Mahidol University
Oregon Health and Science University
Keywords: Medicine
Issue Date: 1-Dec-2004
Citation: Southeast Asian Journal of Tropical Medicine and Public Health. Vol.35, No.4 (2004), 840-844
Abstract: The appearance of drug resistant parasites and the absence of an effective vaccine have resulted in the need for new effective antimalarial drugs. Consequently, a convenient method for in vitro screening of large numbers of antimalarial drug candidates has become apparent. The CyQUANT cell proliferation assay is a highly sensitive fluorescence-based method for quantitation of cell number by measuring the strong fluorescence produced when green GR dye binds to nucleic acids. We have applied the CyQUANT assay method to evaluate the growth of Plasmodium falciparum D6 strain in culture. The GR-nucleic acid fluorescence linearly correlated with percent parasitemia at both 0.75 or 1 percent hematocrit with the same correlation coefficient of r2 = 0.99. The sensitivity of P. falciparum D6 strain to chloroquine and to 3,6-bis-omega-diethylaminoamyloxyxanthone, a novel antimalarial, determined by the CyQUANT assay were comparable to those obtained by the traditional [ 3H]-ethanolamine assay: IC50 value of chloroquine was 54 nM and 51 nM by the CyQUANT and [3H]-ethanolamine assay, respectively; IC50 value for 3,6-bis-omega- diethylaminoamyloxyxanthone was 254 nM and 223 nM by the CyQUANT and [ 3H]-ethanolamine assay, respectively. This procedure requires no radioisotope, uses simple equipment, and is an easy and convenient procedure, with no washing and harvesting steps. Moreover, all procedures can be set up continuously and thus, the CyQUANT assay is suitable in automatic high through-put drug screening of antimalarial drugs.
ISSN: 01251562
Appears in Collections:Scopus 2001-2005

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