Rapid Real-Time PCR Genotyping of Mutations Associated with Sulfadoxine-Pyrimethamine Resistance in Plasmodium falciparum
Open Access
- 1 August 2004
- journal article
- Published by American Society for Microbiology in Antimicrobial Agents and Chemotherapy
- Vol. 48 (8) , 3172-3174
- https://doi.org/10.1128/aac.48.8.2924-2929.2004
Abstract
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.Keywords
This publication has 23 references indexed in Scilit:
- Determinants of Treatment Response to Sulfadoxine-Pyrimethamine and Subsequent Transmission Potential in Falciparum MalariaAmerican Journal of Epidemiology, 2002
- Epidemiology of drug-resistant malariaPublished by Elsevier ,2002
- Minor Groove Binder-Conjugated DNA Probes for Quantitative DNA Detection by Hybridization-Triggered FluorescenceBioTechniques, 2002
- New Real-Time PCR Able To Detect in a Single Tube Multiple Rifampin Resistance Mutations and High-Level Isoniazid Resistance Mutations in Mycobacterium tuberculosisJournal of Clinical Microbiology, 2002
- Rapid Real-Time PCR for Determination of Penicillin Susceptibility in Pneumococcal Meningitis, Including Culture-Negative CasesJournal of Clinical Microbiology, 2002
- Plasmodium falciparum: Underestimation of Dihydrofolate Reductase and Dihydropteroate Synthase Polymorphism in Field Samples: A Technical Shortcoming of Nested PCR Assays with Mutation-Specific PrimersExperimental Parasitology, 2001
- Towards an Understanding of the Mechanism of Pyrimethamine-Sulfadoxine Resistance in Plasmodium falciparum : Genotyping of Dihydrofolate Reductase and Dihydropteroate Synthase of Kenyan ParasitesAntimicrobial Agents and Chemotherapy, 2000
- 3'-Minor groove binder-DNA probes increase sequence specificity at PCR extension temperaturesNucleic Acids Research, 2000
- Multiplex detection of four pathogenic retroviruses using molecular beaconsProceedings of the National Academy of Sciences, 1999
- Allelic discrimination using fluorogenic probes and the 5′ nuclease assayGenetic Analysis: Biomolecular Engineering, 1999