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Antimicrobial Agents and Chemotherapy, May 2004, p. 1803-1806, Vol. 48, No. 5
0066-4804/04/$08.00+0     DOI: 10.1128/AAC.48.5.1803-1806.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Simple and Inexpensive Fluorescence-Based Technique for High-Throughput Antimalarial Drug Screening

Martin Smilkstein,1,2* Nongluk Sriwilaijaroen,3 Jane Xu Kelly,1 Prapon Wilairat,3 and Michael Riscoe1,2

Medical Research Service, Department of Veterans Affairs Medical Center,1 Oregon Health and Science University, Portland, Oregon,2 Department of Biochemistry, Mahidol University, Bangkok, Thailand3

Received 15 August 2003/ Returned for modification 27 October 2003/ Accepted 12 January 2004

Radioisotopic assays involve expense, multistep protocols, equipment, and radioactivity safety requirements which are problematic in high-throughput drug testing. This study reports an alternative, simple, robust, inexpensive, one-step fluorescence assay for use in antimalarial drug screening. Parasite growth is determined by using SYBR Green I, a dye with marked fluorescence enhancement upon contact with Plasmodium DNA. A side-by-side comparison of this fluorescence assay and a standard radioisotopic method was performed by testing known antimalarial agents against Plasmodium falciparum strain D6. Both assay methods were used to determine the effective concentration of drug that resulted in a 50% reduction in the observed counts (EC50) after 48 h of parasite growth in the presence of each drug. The EC50s of chloroquine, quinine, mefloquine, artemisinin, and 3,6-bis-{varepsilon}-(N,N-diethylamino)-amyloxyxanthone were similar or identical by both techniques. The results obtained with this new fluorescence assay suggest that it may be an ideal method for high-throughput antimalarial drug screening.

* Corresponding author. Mailing address: Medical Research Service, Department of Veterans Affairs Medical Center, RD-33, 3710 SW US Veterans Hospital Rd., Portland, OR 97201. Phone: (503) 220-8262, ext. 54131. Fax: (503) 721-1084. E-mail: smilkste{at}earthlink.net.

Antimicrobial Agents and Chemotherapy, May 2004, p. 1803-1806, Vol. 48, No. 5
0066-4804/04/$08.00+0     DOI: 10.1128/AAC.48.5.1803-1806.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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