Spatio-temporal distribution of antimalarial drug resistant gene mutations in a Plasmodium falciparum parasite population from Kilifi, Kenya: A 25-year retrospective study

Abstract

<ns7:p> <ns7:bold>Background:</ns7:bold> Antimalarial drug resistance is a major obstacle to sustainable malaria control. Here we use amplicon sequencing to describe molecular markers of drug resistance in <ns7:italic>Plasmodium falciparum</ns7:italic> parasites from Kilifi county in the coastal region of Kenya over a 25-year period. </ns7:p> <ns7:p> <ns7:bold>Methods:</ns7:bold> We performed <ns7:italic>P. falciparum</ns7:italic> amplicon sequencing on 1162 malaria-infected blood samples collected between 1994 and 2018 to identify markers of antimalarial drug resistance in the <ns7:italic>Pfcrt</ns7:italic> , <ns7:italic>Pfdhfr</ns7:italic> , <ns7:italic>Pfdhp</ns7:italic> s, <ns7:italic>Pfmdr1</ns7:italic> , <ns7:italic>Pfexo</ns7:italic> , <ns7:italic>Pfkelch13</ns7:italic> , <ns7:italic>plasmepsin</ns7:italic> 2/3, <ns7:italic>Pfarps10, Pffd,</ns7:italic> and <ns7:italic>Pfmdr2</ns7:italic> genes. We further interrogated parasite population structure using a genetic barcode of 101 drug resistance-unrelated single nucleotide polymorphisms (SNPs) distributed across the genomes of 1245 <ns7:italic>P. falciparum</ns7:italic> parasites. </ns7:p> <ns7:p> <ns7:bold>Results:</ns7:bold> Two major changes occurred in the parasite population over the 25 years studied. In 1994, approximately 75% of parasites carried the marker of chloroquine resistance, CVIET. This increased to 100% in 1999 and then declined steadily, reaching 6.7% in 2018. Conversely, the quintuple mutation form of sulfadoxine-pyrimethamine resistance increased from 16.7% in 1994 to 83.6% in 2018. Several non-synonymous mutations were identified in the <ns7:italic>Kelch13</ns7:italic> gene, although none of them are currently associated with artemisinin resistance. We observed a temporal increase in the <ns7:italic>Pfmdr1</ns7:italic> NFD haplotype associated with lumefantrine resistance, but observed no evidence of piperaquine resistance. SNPs in other parts of the genome showed no significant temporal changes despite the marked changes in drug resistance loci over this period. </ns7:p> <ns7:p> <ns7:bold>Conclusions:</ns7:bold> We identified substantial changes in molecular markers of <ns7:italic>P. falciparum</ns7:italic> drug resistance over 25 years in coastal Kenya, but no associated changes in the parasite population structure. </ns7:p>

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