Pyrimethamine and proguanil resistance-conferring mutations in Plasmodium falciparum dihydrofolate reductase: polymerase chain reaction methods for surveillance in Africa.


Journal Article

As chloroquine resistance spreads across Africa, the dihydrofolate reductase (DHFR) inhibitors pyrimethamine and proguanil are being used as alternative first-line drugs for the treatment and prevention of Plasmodium falciparum malaria. Resistance to these drugs is conferred by point mutations in parasite DHFR. These point mutations can be detected by polymerase chain reaction (PCR) assays, but better methods for sample collection, DNA extraction, and a diagnostic PCR are needed to make these assays useful in malaria-endemic areas. Here we report methods for collecting fingerstick blood onto filter paper strips that are air-dried, then stored and transported at room temperature. Cell lysis and DNA extraction are accomplished by boiling in Chelex-100. We also report a nested PCR technique that has improved sensitivity and specificity. These procedures readily detect mixed infections of parasites with both sensitive and resistant genotypes (confirmed by direct sequencing) and are reliable at parasite densities less than 250/mm3 in field surveys.

Full Text

Duke Authors

Cited Authors

  • Plowe, CV; Djimde, A; Bouare, M; Doumbo, O; Wellems, TE

Published Date

  • June 1995

Published In

Volume / Issue

  • 52 / 6

Start / End Page

  • 565 - 568

PubMed ID

  • 7611566

Pubmed Central ID

  • 7611566

International Standard Serial Number (ISSN)

  • 0002-9637

Digital Object Identifier (DOI)

  • 10.4269/ajtmh.1995.52.565


  • eng

Conference Location

  • United States