Table 1.
P. falciparum and P. vivax K13 propeller domain primers.
Primary and secondary PCR primers for each species and respective annealing temperatures are shown.
Fig 1.
Schematic representation of the P. falciparum K13 gene.
(A) Schematic depicting the P. falciparum K13 gene size and propeller domain region. A 3D model is shown for the predicted propeller domain. (B) Species specific nested PCR workflow for amplifying the K13 gene and propeller domain. The protein structure of the Keap1 protein (Protein Data Bank: 2Z32) was used to generate the 3D model of the P. falciparum K13 propellor domain.
Fig 2.
Alignment of kelch gene for P. falciparum and P. vivax.
Nucleotide and amino acid sequence alignment of the kelch gene for P. falciparum and P. vivax; disagreements in nucleotide (black) and amino acid (color) are highlighted. The yellow annotation above the sequence indicates the gene region that encodes the kelch propeller domain; blue squares over the P. falciparum sequence denotes K13 propeller mutations reported by Ariey et al [14]. Forward primers are annotated in dark green and reverse primers in light green. Shown are sequences for the isolates 3D7 (P. falciparum) and Sal-1 (P. vivax).
Table 2.
Genetic distance matrix of kelch propeller domain of various Plasmodium species.
Amino acid and nucleotide sequence percent (%) identify (e.g. percent of residues that are identical) are shown in the table. Amino acid similarity is shown in bold numbers and nucleotide similarity in italicized numbers.
Table 3.
P. falciparum non-synonymous amino acid changes in the kelch propeller domain as compared to P. vivax.
Relative to alignment between P. falciparum 3D7 and P. vivax Sal-I.
Fig 3.
P. falciparum K13 gene species-specific amplification.
The Plasmodium falciparum species specific primers were tested on multiple Plasmodium species, including human, rodent, and non-human primate malaria parasites from various geographical regions. The expected K13 propeller domain PCR product for P. falciparum is 784 base pairs. Panels (A) and (B) show human malaria parasites tested; Panel (C) shows non-human primate and rodent malaria parasites tested. Blue text denotes samples that were amplified by the PCR protocol.
Fig 4.
P. vivax K12 gene species-specific amplification.
The Plasmodium vivax species specific primers were tested on multiple Plasmodium species, including human, rodent, and non-human primate malaria parasites from various geographical regions. The expected K12 propeller domain PCR product for P. vivax is 792 base pairs. Panels (A) and (B) show human malaria parasites tested; Panel (C) shows non-human primate and rodent malaria parasites tested. Blue text denotes samples that were amplified by the PCR protocol.