Novel pfk13 Gene Mutations Are Potentially Associated with Artemisinin Resistance in Plasmodium falciparum : A Clinical–Genetic Study

Background: Artemisinin-based combination therapies remain the first-line treatment for Plasmodium falciparum malaria. However, emerging resistance, often linked to mutations in the Kelch 13 ( pfk13 ) gene, poses a substantial threat to malaria control efforts. This study investigated the molecular profile of pfk13 in P. falciparum isolates from a health facility in Warri, South-South Nigeria. Methods: A total of 284 patients were screened using rapid diagnostic tests. Genomic DNA was extracted from 50 confirmed cases using Whatman filter paper-collected samples. The pfk13 gene was amplified through nested polymerase chain reaction, sequenced, and analyzed using Qiagen CLC Genomics Workbench (v23.0.1). The sequences were aligned to those of the reference P. falciparum 3D7 strain (PF3D7_1343700), and the translated amino acid sequences were subsequently examined for mutations. Results: Among the 50 isolates, 43 nonsynonymous mutations were identified, including 16 novel and previously unreported variants: E9K, K10N4, K15V, K17R, A21K, S23I, I24E, S25K, F27I, S28V, N40K, D42K, D43K, D44R, S45Y, and G46Y. None of the validated pfk13 mutations known to confer artemisinin resistance were detected. Conclusion: Although no validated resistance-associated mutations were detected, the identification of multiple novel nonsynonymous variants may reflect the ongoing regional evolutionary dynamics of the pfk13 gene in P. falciparum . These findings underscore the need for functional and phenotypic studies to elucidate the potential role of these variants in modulating artemisinin susceptibility.