Pathogen genotyping in polyclonal infections: application of a fluorogenic polymerase-chain-reaction assay in malaria

J Infect Dis. 2003 Oct 15;188(8):1245-9. doi: 10.1086/378521. Epub 2003 Oct 6.

Abstract

Pathogen genotyping of polyclonal infections is limited by 2 major drawbacks: (1). how to establish whether multiple mutations detected in 1 gene belong to the same clone and (2). how to evaluate the proportion of different genotypes in a given sample. For drug-resistance genotyping in Plasmodium falciparum malaria, we address these problems by using a fluorogenic assay that combines fluorescence-resonance energy transfer, between fluorophores present on a probe and a polymerase-chain-reaction primer, and a melt-curve analysis. We demonstrate that this tool allows a more accurate insight into the P. falciparum populations present in complex biological samples.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA, Protozoan / analysis
  • Fluorescence Resonance Energy Transfer
  • Fluorescent Dyes*
  • Genotype
  • Humans
  • Malaria, Falciparum / parasitology*
  • Plasmodium falciparum / classification*
  • Plasmodium falciparum / genetics
  • Polymerase Chain Reaction / methods*
  • Tetrahydrofolate Dehydrogenase / genetics

Substances

  • DNA, Protozoan
  • Fluorescent Dyes
  • Tetrahydrofolate Dehydrogenase