Plasmodium vivax in the Democratic Republic of East Timor: Parasite prevalence and antifolate resistance-associated mutations

In the Democratic Republic of East Timor, Plasmodium falciparum and Plasmodium vivax malaria coexist, but limited information is available about the latter species. Consequently, the prevalence of P. vivax and of its corresponding antifolate resistance-associated mutations in the pvdhfr and pvdhps genes was assessed here. Blood samples were collected from 650 individuals distributed among six districts, over two different periods, by either passive case detection (PCD) or active case detection (ACD). As expected, malaria was over-represented in the PCD sample (26% PCD vs 5% ACD), because the infection increases medical care seeking. Additionally, the relative frequency of P. vivax infections in symptomatic individuals (37%) was twice as high as the one in the asymptomatic sampling group (18%), suggesting that that this parasite is accounting for a significant proportion malaria-attributed morbidity. The frequency of specific sulfadoxine-pyrimethamine resistance-associated mutations genes was ascertained in P. vivax positive samples by PCR-RFLP. Although no mutants were detected in codons 383 and 553 of pvdhps, 48%, 76% and 82% of P. vivax-infected samples harbored the dhfr 33L, 58R and 117N mutations, respectively. Additionally, the frequency of parasites carrying both pvdhfr 58R and 117N mutant alleles accounted for a third of all genotypes analyzed, most likely due to inadvertent SP use in the past. In conclusion, evidence-based information is provided to promote optimized drug deployment and limit the evolution of resistance to antifolate resistance in P. vivax from East Timor.