Multiplex PCR–RFLP methods for pfcrt, pfmdr1 and pfdhfr mutations in Plasmodium falciparum

Plasmodium falciparum drug resistance is a major factor for the death toll of malaria. Resistance has been associated with specific single nucleotide polymorphisms (SNPs) in the parasite genes pfmdr1 (N86Y) and pfcrt (K76T) associated with quionoline antimalarial resistance, and pfdhfr (N51I, C59R, S108N) correlated with resistance of the antifolate combination sulfadoxine–pyrimethamine. These SNPs constitute the basis for the surveillance of drug resistance through high sensitive molecular methods in malaria endemic countries. In this work, we developed a multiplex PCR–RFLP protocols for the diagnosis of these molecular markers, leading to significant decreases in reagent costs, time, number of manipulations and hence human resources.