Plasmodium falciparum gene mutations that confer resistance to Artemether-Lumefantrine at Kasangati Health Center IV Wakiso District-Uganda

Abstract

Background: Artemisinin-based combination therapy (ACT) is the mainstay of efforts for treating Plasmodium falciparum malaria. Artemisinin and its derivatives are the most potent and rapidly acting anti-malarials. Artemether-Lumefrantrine (AL) is one of the ACTs recommended by Uganda National Malaria Control Programme (UNMCP) to be used as the first line therapy to treat uncomplicated malaria in Uganda.

Aim: To determine the prevalence of P. falciparum gene mutations S769N in PfATPase6 gene and N86Y in pfmdr-1 gene associated with resistance to artemether-lumefantrine. Method: This was a cross sectional study where a total of 85 archived blood samples stored at 80oC were used. This archived samples were microscopically and PCR positive for P. falciparum. Parasite DNA was extracted from whole blood samples, using Qiagen Method, Nested PCR and Allele-Specific Restriction Enzyme Digestion was applied to detect gene mutations at positions S769N and N86Y on the pf ATPase 6 and pf mdr-1 gene, respectively.

Results: Out the 85 samples, 36 (42.4%) samples amplified successfully and tested for gene mutations at position S769N mutation on the pf ATPASE 6. No genetic mutations were detected in any of the 36 samples; only the wild type genes were present. For the N86Y mutation on the pfMDR-1 gene, 63 (74.1%) samples amplified, of these 2 (3.2%) samples were of the mutant type while 61 (96.8%) samples were of the wild type gene. Conclusion: The results indicate that the S769N mutation on the PfATPase 6 gene that confers resistance to artemisinin may not yet be present in P. falciparum isolates at Kasangati Health Centre IV, in Uganda and so AL use should be continued. The presence of only 3.2% mutation at position N86Y on pfmdr-1 gene that confers resistance to quinolines gives hope that several antimalarials have re-gained usefulness in the country.

Key: Artemisinin, pf ATPase, pf mdr-1, S769N mutation, N86Y mutation, Anti-Malarial Drug Resistance