Molecular Surveillance Of Antimalarial Drug Resistance Genes In Plasmodium Falciparum Isolates In Indonesia

Abstract

Malaria is a disease caused by infection of the Plasmodium parasite that spreads through Anopheles mosquitoes, which mainly plagues humans living in regions with poor healthcare and countries around the equator. Among the five species capable of causing malaria in humans, Plasmodium falciparum and Plasmodium vivax are noted as the most threatening species, as their manifestations in humans are much stronger compared to the other species. Although various antimalarial treatments have been given to stricken patients, cases of resistances to treatments rises in numerous places in the world. The three most notable genes where the mutation related to drug resistance commonly occurs are the PfCRT, PfMDR1, and PfKelch13. Existing diagnostic methods can only detect the presence of malaria parasites in a contracted human, but not mutations related to drug resistance. Employing sample amplification using PCR followed by Oxford Nanopore Technologies (ONT) sequencing utilizing the Rapid Barcoding Kit (SQK-RBK110.96), this study seeks to develop and optimize a protocol to profile genetic variances of Plasmodium falciparum collected from clinical patients in Papua, Indonesia. Although optimization has yet to produce persistent results among trials, optimization attempts had produced potential amplification configurations which can aid future studies to develop an established protocol for amplification of the whole gene of interest aforementioned, including adjustments in annealing temperature, extension duration, and extension temperature.