Prevalence of mutations in Plasmodium falciparum genes associated with resistance to different antimalarial drugs in Nyando, Kisumu County in Kenya

Infect Genet Evol. 2020 Mar:78:104121. doi: 10.1016/j.meegid.2019.104121. Epub 2019 Nov 19.

Abstract

Resistance to the mainstay antimalarial drugs is a major concern in the control of malaria. Delayed Plasmodium falciparum parasite clearance has been associated with Single Nucleotide Polymorphisms (SNPs) in the kelch propeller region (K13). However, SNPs in the Pf-adaptor protein complex 2 mu subunit (Pfap2-mu), Pfcrt and Pfmdr1 are possible markers associated with multi-drug resistance. Here, we explored the prevalence of SNPs in the K13, Pfap2-mu, Pfcrt, and Pfmdr1 in 94 dried blood spot field isolates collected from children aged below 12 years infected with P. falciparum during a cross-sectional study. The samples were collected in 2015 during the peak malaria transmission season in the Nyando region of Western Kenya before treatment with Artemether-Lumefantrine, the first-line artemisinin-based combination therapy (ACT) in Kenya. However, 47 of the 94 samples had recurrent parasitemia and were interrogated for the presence of the SNPs in K13 and Pfap2-mu. We used PCR amplification and sequencing to evaluate specific regions of K13 (codons 432-702), Pfap2-mu (codons 1-350), Pfmdr1 (codons 86, 1034-1246), and Pfcrt (codons 72-76) gene(s). The majority of parasites harbored the wild type K13 sequence. However, we found a unique non-synonymous W611S change. In silico studies on the impact of the W611S predicted structural changes in the overall topology of the K13 protein. Of the 47 samples analyzed for SNPs in the Pfap2-mu gene, 14 (29%) had S160 N/T mutation. The CVIET haplotype associated with CQ resistance in the Pfcrt yielded a 7.44% (7/94), while CVMNK haplotype was at 92.56%. Mutations in the Pfmdr1 region were detected only in three samples (3/94; 3.19%) at codon D1246Y. Our data suggest that parasites in the western part of Kenya harbor the wildtype strains. However, the detection of the unique SNP in K13 and Pfap2-mu linked with ACT delayed parasite clearance may suggest slow filtering of ACT-resistant parasites.

Keywords: ACT; Delayed clearance; PfK13; Pfap2mu; Plasmodium falciparum; SNPs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP Binding Cassette Transporter, Subfamily B, Member 1 / genetics
  • Antimalarials / pharmacology*
  • Antimalarials / therapeutic use
  • Artemisinins / therapeutic use
  • Child
  • Child, Preschool
  • Drug Resistance, Microbial / genetics*
  • Humans
  • Infant
  • Kenya / epidemiology
  • Malaria, Falciparum / epidemiology
  • Malaria, Falciparum / parasitology*
  • Mutation*
  • Plasmodium falciparum / drug effects
  • Plasmodium falciparum / genetics*
  • Plasmodium falciparum / isolation & purification
  • Polymorphism, Single Nucleotide
  • Prevalence
  • Protozoan Proteins / chemistry
  • Protozoan Proteins / genetics

Substances

  • ATP Binding Cassette Transporter, Subfamily B, Member 1
  • Antimalarials
  • Artemisinins
  • Protozoan Proteins
  • artemisinin