Allele-specific, nested, one tube PCR: application to Pfmdr1 polymorphisms in Plasmodium falciparum

An allele-specific, one tube PCR for the sensitive and reliable detection of point mutations in Plasmodium falciparum DNA is described. Design of specific internal primers and optimization of the PCR is simple, and the procedure is robust and sensitive. Single nucleotide polymorphisms at codons 184, 1034, 1042 and 1246 of the P, falciparum multidrug resistance gene Pfmdr1, were examined in 6 laboratory isolates, to validate the technique.