Common origin and fixation of Plasmodium falciparum dhfr and dhps mutations associated with sulfadoxine-pyrimethamine resistance in a low-transmission area in South America

Abstract

Recent studies indicated that sensitive parasites could increase in frequency in a population when drugs are removed, suggesting that the life span of affordable antimalarial drugs could be expanded. We studied 97 samples from Bolivar State, Venezuela, an area where sulfadoxine-pyrimethamine (SP) has not been used for 8 years due to its ineffectiveness. We characterized point mutations in two genes that have been implicated in resistance to SP, dihydrofolate reductase (dhfr) and dihydropteroate synthase (dhps). We also assayed neutral microsatellite markers around the dhfr (chromosome 4) and dhps (chromosome 8) genes and on chromosomes 2 and 3 to track the origin and spread of resistant alleles. We found that drug-resistant SP mutants are fixed in the population. Two genotypes were present in the samples, dhfr(50R/51I/108N) dhps(437G/540E/581G) (90.7%) and dhfr(51I/108N) dhps(437G/581G) (9.3%). We show a single microsatellite haplotype for all of the dhfr and dhps alleles, and the alleles at the microsatellite loci are different from those present in Africa. Thus, in these samples from Venezuela, there is a single origin for both dhfr and dhps SP-resistant alleles, and these alleles originated independently of those characterized from Africa. Furthermore, this is the first report of a "hitchhiking effect" on the genetic variation around dhps due to selection by SP using an extensive set of microsatellite markers. Our results indicate that, in areas where there is limited gene flow, the fixation of drug-resistant parasites in the population is stable, even after drug selection is relaxed.

FIG. 1.

Graphical display of H_e ±1 standard deviation across chromosome 4 around dhfr. Shown are the entire region assayed with microsatellite markers (A) and a close-up of the smaller region of the noted low heterozygosity (B). The dashed line in each graph is the mean H_e calculated from loci on chromosomes 2 and 3. On the x axis, negative numbers are positions 5′ to the gene and positive numbers are positions 3′ to the gene. The error bars show 1 standard deviation.

FIG. 2.

Graphical display of H_e (±1 standard deviation) across chromosome 8 around dhps. Shown are the entire region assayed with microsatellite markers (A) and a close-up of the smaller region of the noted low heterozygosity (B). The dashed line in each graph is the mean H_e calculated from loci on chromosomes 2 and 3. On the x axis, negative numbers are positions 5′ to the gene and positive numbers are positions 3′ to the gene. The error bars show 1 standard deviation.