Detection of mutations associated with artemisinin resistance in K13 gene of Plasmodium falciparum parasites.

Abstract

Objective: this study aims to develop a methodology for detecting mutations in the K13 gene and determine mutation frequencies in clinical samples. Method: total DNAs were extracted blood samples collected from 50 patients in failure with artemisinin treatment in the Central Highlands region. A fragment of the K13 gene was amplified, purified, and sequenced by the Sanger method. The K13 sequences were analysed by using Bioedit and aligned with the reference sequence to determine K13 mutations. Results: successfully amplified the K13 gene segment with size 799 bp in P. falciparum. After sequencing, there was a nucleotide substitution of A>G at position 1740, leading to changes in amino acids C>Y at the respective position 580. In the 50 patient samples, 41/50 (82%) samples showed the C580Y mutation, 9/50 (18%) of the samples had no mutation, and there was no other mutation. Conclusion: the authors have successfully developed and optimised a procedure for detecting the mutation C580Y in K13 in P. falciparum and determined the mutation frequency in the K13 gene.