Abstract
Background: Plasmodium falciparum (Pf) resistance to antimalarial drugs is a major impediment to malaria control. The Pf.Kelch 13 (PfK13) gene has been largely reported to be associated with artemisinin resistance. However, recent studies have shown artemisinin resistance without Kech13 mutations suggesting the implication of others genes in artemisinin resistance. In this current study, we focused on mutations in Pf.actin-binding protein coronin, Pf.cysteine desulfurase and Pf.plasmepsin 2 gene, three putative candidates recently were reported to be involved in artemisinin, lumefantrine and piperaquine resistance respectively. Method: Archived blood samples previously collected from asymptomatic school children from December 2016 to October 2018 were used in this study. Genomic DNA was extracted using ISOLATE II Genomic DNA kit. After PCR amplification, amplicons were purified and sequenced by capillary sequencing. Reads were analyzed for the identification of point mutations previously reported to be involved in drug selection. Results: Mutations R100K, and G50E involved in reduced artemisinin susceptibility were detected in Pfcoronin. From 2016/17 to 2018 the allele 100k increased frequency (11.2%); while 50E was only observed in 2018 time point reaching 11.1%. Lumefantrine selection marker K65, in codon (K65Q) was observed at 14.2% in Pfcysteine desulfurase, and the mutant’ allele 65Q gradually increased frequency from 28.5% in 2016/17 to 57.1% in 2018. Pf.plasmepsin 2 was the less polymorphic gene. Several other polymorphism codons and single nucleotide variants were detected. Conclusion: The findings indicate the presence of mutations associated with reduced artemisinin susceptibility and lumefantrine selection marker. Therefore, the results call for continuous monitoring of molecular makers in Mbita parasites.
Highlights
Plasmodium falciparum (Pf) resistance to antimalarial drugs is a major impediment to malaria control
We focused on mutations in Pf.actin-binding protein coronin, Pf.cysteine desulfurase and Pf.plasmepsin 2 gene, three putative candidates recently were reported to be involved in artemisinin, lumefantrine and piperaquine resistance respectively
The following five codons (G50E, S81S, I68K, Q61P, and N60N) showed no variation in 2016/17 whereas mutants were observed at frequencies of 11.1% in the 2018 time point
Summary
Plasmodium falciparum (Pf) resistance to antimalarial drugs is a major impediment to malaria control. In Kenya, malaria is the major cause of morbidity and mortality ordinarily affecting children under 5 years of age and pregnant women, accounting for 19% of outpatient attendance to health centres [4]. Clinical cases are estimated at around 6.7 million each year, with 4000 deaths occurring among children, making malaria a major threat in Kenya [5]. The use of ACTs has been successful and has contributed to reducing malaria burden This success is threatened by the emergence and spread of Plasmodium falciparum artemisinin-resistant parasites, first reported in Cambodia in 2007 and has subsequently spread over Greater Mekong Subregion (GMS) [13] [14] [15], where the former
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