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Antimicrobial Agents and Chemotherapy, August 2004, p. 2924-2929, Vol. 48, No. 8
0066-4804/04/$08.00+0     DOI: 10.1128/AAC.48.8.2924-2929.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Rapid Real-Time PCR Genotyping of Mutations Associated with Sulfadoxine-Pyrimethamine Resistance in Plasmodium falciparum

Department of Epidemiology, School of Public Health,¹ Department of Microbiology and Immunology, School of Medicine, University of North Carolina, Chapel Hill, North Carolina,⁴ Department of Community Health, College of Medicine, Blantyre, Malawi,² Department of Epidemiology, University of Michigan, Ann Arbor, Michigan³

Received 30 January 2004/ Returned for modification 10 March 2004/ Accepted 19 April 2004

The resistance of Plasmodium falciparum to sulfadoxine-pyrimethamine (SP) is an emerging public health threat. Resistance to these drugs is associated with point mutations in the genes encoding dihydropteroate synthase (DHPS) and dihydrofolate reductase (DHFR). We describe here an assay using real-time PCR and sequence-specific probes that detects these mutations. Using DNA from plasmids, cultured strains, and clinical samples, real-time PCR could distinguish four DHPS polymorphisms (codons 437, 540, 581, and 613) and three DHFR polymorphisms (codons 51, 59, and 108). This assay is rapid and sensitive, with a detection limit of 10 copies in most cases. This assay is amenable to large-scale studies of drug resistance.

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