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A NOVEL DNA-BASED MICROFLUORIMETRIC METHOD TO EVALUATE ANTIMALARIAL DRUG ACTIVITY

This paper describes the development of a novel microfluorimetric assay to measure the inhibition of Plasmodium falciparum based on the detection of parasitic DNA by intercalation with PicoGreen^®. The method was used to determine parasite inhibition profiles and 50% inhibitory concentration values of known or potential antimalarial drugs. Values for parasite inhibition with known antimalarial drugs using the PicoGreen^® assay were comparable with those determined by the standard method based upon the uptake of ³H-hypoxanthine and the Giemsa stain microscopic technique. The PicoGreen^® assay is rapid, sensitive, reproducible, easily interpreted, and ideally suited for screening of large numbers of samples for antimalarial drug development.

Received April 4, 2003. Accepted for publication October 1, 2003.

Acknowledgments: Special thanks are given to Phil Rosenthal, Dennis Kyle, and Jeff Ryan for their continuous and generous support. We also thank all members of the laboratory of Eduardo Ortega-Barria for helpful discussions and encouragement.

Financial support: This work was supported by the International Cooperative Biodiversity Groups Program, award #1U01 TW01021-01. The laboratory of Eduardo Ortega-Barria is partially supported by National Institutes of Health grant 1R03 TW01076.

Authors’ addresses: Yolanda Corbett, Liuris Herrera, Jose Gonzalez, Luz I. Romero, and Eduardo Ortega-Barría, Instituto de Investigaciones Científicas Avanzadas y Servicios de Alta Tecnología, Ciudad del Saber, PO Box 7250, Zona 5, Clayton, Panama City, Panama. Luis Cubilla, Laboratorio de Productos Naturales, Universidad de Panama, Panama City, Panama. Todd L. Capson, Smithsonian Tropical Research Institute, Apartado 2072, Balboa, Ancon, Panama. Phyllis D. Coley and Thomas Kursar, Department of Biology, University of Utah, Salt Lake City, UT 84112 and Smithsonian Tropical Research Institute, Apartado 2072, Balboa, Ancon, Panama.

Reprint requests: Eduardo Ortega-Barría, Instituto de Investigaciones Científicas Avanzadas y Servicios de Alta Tecnología, Ciudad del Saber, PO Box 7250, Zona 5, Clayton, Panama City, Panama, Telephone: 507-317-0012, Fax: 507-317-0023, E-mail: eortega{at}senacyt.gob.pa.

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