title: Point mutations in the dihydrofolate reductase and dihydropteroate synthetase genes and in vitro susceptibility to pyrimethamine and cycloguanil of Plasmodium falciparum isolates from Papua New Guinea.
abstract: Plasmodium falciparum isolates from 24 Papua New Guinean patients with symptomatic malaria were tested for susceptibility to pyrimethamine and cycloguanil. Thirteen isolates were sensitive to both agents and the remainder exhibited varying degrees of resistance. No isolates were found to be resistant to one agent yet sensitive to the other and a positive correlation suggesting cross-resistance was found. Parasite DNA extracted from the patients' stained blood slides was amplified and sequenced to examine point mutations in the dihydrofolate reductase (DHFR) and dihydropteroate synthetase genes (DHPS) associated with antifolate resistance. All resistant isolates possessed mutations in the DHFR gene at codon 108, the majority changing from Ser to Asn, but one isolate from Ser to Thr, a change not previously reported in field isolates. A second mutation of the DHFR gene at Cys-59 to Arg was present in isolates with higher level resistance, but not exclusively so. Sequencing the DHPS gene, as a predictor of sulfadoxine resistance, revealed only one example that was different from DHPS alleles of sensitive isolates.
authors.affiliation: Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia.
authors.forename: J C
authors.initials: JC
authors.lastname: Reeder
authors.forename: K H
authors.initials: KH
authors.lastname: Rieckmann
authors.forename: B
authors.initials: B
authors.lastname: Genton
authors.forename: K
authors.initials: K
authors.lastname: Lorry
authors.forename: B
authors.initials: B
authors.lastname: Wines
authors.forename: A F
authors.initials: AF
authors.lastname: Cowman
mesh_terms: D000818:Animals; D000962:Antimalarials / Q000494:pharmacology*; D001483:Base Sequence; D017931:DNA Primers / Q000737:chemistry; D004094:Dihydropteroate Synthase / Q000235:genetics*; D004351:Drug Resistance / Q000235:genetics; D005493:Folic Acid Antagonists / Q000494:pharmacology*; D006801:Humans; D016778:Malaria, Falciparum / Q000469:parasitology; D008969:Molecular Sequence Data; D010963:Plasmodium falciparum / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics*; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D002727:Proguanil; D011739:Pyrimethamine / Q000494:pharmacology; D017422:Sequence Analysis, DNA; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics*; D014227:Triazines / Q000494:pharmacology
chemical_list: D000962:Antimalarials; D017931:DNA Primers; D005493:Folic Acid Antagonists; D014227:Triazines; C026009:cycloguanil; D013762:Tetrahydrofolate Dehydrogenase; D004094:Dihydropteroate Synthase; D002727:Proguanil; D011739:Pyrimethamine