A new method for detection of pfmdr1 mutations in Plasmodium falciparum DNA using real-time PCR

Malar J. 2004 May 7:3:9. doi: 10.1186/1475-2875-3-9.

Abstract

Background: Surveillance for drug-resistant Plasmodium falciparum should be a component of malaria control programmes. Real-time PCR methods for the detection of parasite single-nucleotide polymorphisms (SNPs) and gene amplification could be useful survellance tools.

Methods: A real-time PCR assay has been developed that identifies single nucleotide polymorphisms (SNPs) at amino acids 86, 184, 1034 and 1042 in the P. falciparum multi-drug resistant (pfmdr 1) gene that may be associated with anti-malarial drug resistance.

Results: This assay has a sensitivity and specificity of 94% and 100% when compared to traditional PCR methods for genotyping. Only 54 of 68 (79%) paired pre- and post-culture DNA samples were concordant at all four loci.

Conclusion: Real-time PCR is a sensitive and specific method to detect SNP's in pfmdr 1. Genotypes of parasites after in vitro culture may not reflect that seen in vivo.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • ATP-Binding Cassette Transporters / genetics
  • Animals
  • Antimalarials / pharmacology*
  • Antimalarials / therapeutic use
  • DNA, Protozoan / analysis
  • DNA, Protozoan / classification
  • DNA, Protozoan / genetics
  • Drug Resistance, Multiple / genetics
  • Genes, MDR / genetics*
  • Genotype
  • Humans
  • Malaria, Falciparum / drug therapy
  • Malaria, Falciparum / parasitology*
  • Plasmodium falciparum / classification
  • Plasmodium falciparum / drug effects
  • Plasmodium falciparum / genetics*
  • Polymerase Chain Reaction / methods*
  • Polymerase Chain Reaction / standards
  • Polymorphism, Single Nucleotide / genetics*
  • Protozoan Proteins / genetics
  • Sensitivity and Specificity

Substances

  • ATP-Binding Cassette Transporters
  • Antimalarials
  • DNA, Protozoan
  • Protozoan Proteins
  • mdr gene protein, Plasmodium