DRP004572
Nanopore sequencing of drug-resistance-associated genes in malaria parasites, Plasmodium falciparum
Here we report the application of a portable sequencer, MinION, for genotyping malaria parasite, Plasmodium falciparum, which is particularly designed for the analysis in developing countries. In this design, amplicon mixture of nine representative genes causing resistance for anti-malaria drug is diagnosed. Firstly, we developed the procedure for four laboratory strains (3D7, Dd2, 7G8, and K1) and, then, applied the developed procedure for ten clinical samples. Even though the average base call accuracy of MinION sequencer was 74.3%, >50 reads at a given position yields the accuracy of the SNP call at the precision and recall rate of 0.92 and 0.76, respectively. Long reads of MinION also enabled identifying mutual associations of multiple SNPs or haplotypes. Based on the SNP information, possible status of the drug resistance in each of ten clinical samples was inferred. Taking advantages of MinION sequencing, this approach will change the way of sequencing diagnosis of malaria parasites especially in developing countries.