Rapid detection of point mutations in Plasmodium falciparum genes associated with antimalarial drugs resistance by using High-Resolution Melting analysis

J Microbiol Methods. 2009 Aug;78(2):165-70. doi: 10.1016/j.mimet.2009.05.013. Epub 2009 May 22.

Abstract

We have developed a High-Resolution DNA Melting method to detect mutations related to Plasmodium falciparum resistance. This method is based on real-time PCR followed by High Resolution Melting ramping from 67 degrees C to 80 degrees C with fluorescence data acquisition set at 0.1 degrees C increments. The accuracy of the technique was assessed using 177 P. falciparum clinical isolates and two reference strains. Results perfectly matched those obtained by DNA sequencing for some important genetic markers of P. falciparum resistance. This technique could be of great value for epidemiological studies, especially in developing countries.

Publication types

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

MeSH terms

  • Animals
  • Antimalarials / pharmacology*
  • DNA, Protozoan / genetics*
  • Drug Resistance*
  • Hot Temperature
  • Humans
  • Nucleic Acid Denaturation
  • Parasitic Sensitivity Tests / methods*
  • Plasmodium falciparum / drug effects*
  • Plasmodium falciparum / genetics
  • Point Mutation*
  • Polymerase Chain Reaction / methods
  • Sensitivity and Specificity
  • Transition Temperature*

Substances

  • Antimalarials
  • DNA, Protozoan