Comprehensive analysis of molecular markers linked to antimalarial drug resistance in Plasmodium falciparum in Northern, Northeastern and Eastern Uganda

Abstract Background In Uganda, antimalarial resistance in Plasmodium falciparum poses serious public health and treatment challenges. Globally, recent data have highlighted the roles of following genes in malaria resistance: Plasmodium falciparum dihydrofolate reductase (Pfdhfr), Plasmodium falcipar...

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Main Authors: Peter Olupot-Olupot, George Paasi, Thomas Katairo, Jimmy Patrick Alunyo, Alice Nakiyemba, Gilbert Gilibrays Ocen, Stephen Pande, Florance Alaroker, William Okiror, Emmaluel Ocen, Alex Oula, Charles Benard Okalebo, Ongodia Paul, Denis Amorut, Stephen Tukwasibwe, Susan Nabadda Ndidde, Isaac Sewanyana, Samuel L. Nsobya
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Language:English
Published: BMC 2025-06-01
Series:Malaria Journal
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Online Access:https://doi.org/10.1186/s12936-025-05439-x
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author Peter Olupot-Olupot
George Paasi
Thomas Katairo
Jimmy Patrick Alunyo
Alice Nakiyemba
Gilbert Gilibrays Ocen
Stephen Pande
Florance Alaroker
William Okiror
Emmaluel Ocen
Alex Oula
Charles Benard Okalebo
Ongodia Paul
Denis Amorut
Stephen Tukwasibwe
Susan Nabadda Ndidde
Isaac Sewanyana
Samuel L. Nsobya
author_facet Peter Olupot-Olupot
George Paasi
Thomas Katairo
Jimmy Patrick Alunyo
Alice Nakiyemba
Gilbert Gilibrays Ocen
Stephen Pande
Florance Alaroker
William Okiror
Emmaluel Ocen
Alex Oula
Charles Benard Okalebo
Ongodia Paul
Denis Amorut
Stephen Tukwasibwe
Susan Nabadda Ndidde
Isaac Sewanyana
Samuel L. Nsobya
author_sort Peter Olupot-Olupot
collection DOAJ
description Abstract Background In Uganda, antimalarial resistance in Plasmodium falciparum poses serious public health and treatment challenges. Globally, recent data have highlighted the roles of following genes in malaria resistance: Plasmodium falciparum dihydrofolate reductase (Pfdhfr), Plasmodium falciparum dihydropteroate synthetase (Pfdhps), Plasmodium falciparum chloroquine resistance transporter (Pfcrt), Plasmodium falciparum multidrug resistance gene 1 (Pfmdr1), and Plasmodium falciparum K13 propeller domain (Pfk13). This study investigated the prevalence and characteristics of P. falciparum molecular markers linked to antimalarial resistance in Northern, Northeastern, and Eastern Uganda. Methods This cross-sectional study collected 200 dried blood samples from children (2 months to 12 years) in Northern, Eastern, and Northeastern Uganda. Samples were from malaria-positive cases confirmed by rapid diagnostic tests and microscopy. Genomic DNA was extracted from these samples and analysed using Molecular Inversion Probes to detect Plasmodium falciparum genetic mutations. The sequencing was performed on the Illumina MiSeq platform, and raw data was organized and analysed with MIPTools software. Results The study sequenced over 50% of the samples at each site as follows: Apac 87.7% (43/49), Moroto 68.0% (34/50), Soroti 65.0% (13/20) and Mbale 53.1% (43/81). The Pfk13 A675V and C469Y mutations varied from 0 to 23.3% and 8.3–14.3%, in four sites, with consistently low prevalence in Apac. The Pfdhfr N51I and S108N mutations were fixed in all districts, while C59R was fixed in Moroto and nearing fixation (92–97%) in other regions. The emerging I164L mutation ranged from 1 to 10% in all sites. The Pfdhps A437G and K540E mutations were fixed in Soroti, with 3–5% wild-type prevalence in other sites. The A581G mutation showed 2.3% mixed genotypes in Mbale only. The Pfcrt K76T was predominantly wild type, except for 5% mutants in Mbale and Moroto. The pfmdr1 N86Y were wild type across all districts, except for 15% mixed genotypes in Soroti. Conclusion This study reveal rising partial artemisinin resistance and widespread antifolate resistance surpassing WHO thresholds in Northern, Northeastern, and Eastern Uganda. Emerging super-resistant parasites pose a serious threat to malaria control, necessitating urgent enhanced surveillance and alternative treatment strategies.
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spelling doaj-art-9ca2c7d22b4e45f8ae22308d0f482cf82025-08-20T02:39:45ZengBMCMalaria Journal1475-28752025-06-012411910.1186/s12936-025-05439-xComprehensive analysis of molecular markers linked to antimalarial drug resistance in Plasmodium falciparum in Northern, Northeastern and Eastern UgandaPeter Olupot-Olupot0George Paasi1Thomas Katairo2Jimmy Patrick Alunyo3Alice Nakiyemba4Gilbert Gilibrays Ocen5Stephen Pande6Florance Alaroker7William Okiror8Emmaluel Ocen9Alex Oula10Charles Benard Okalebo11Ongodia Paul12Denis Amorut13Stephen Tukwasibwe14Susan Nabadda Ndidde15Isaac Sewanyana16Samuel L. Nsobya17Mbale Clinical Research Institute (MCRI)Mbale Clinical Research Institute (MCRI)Infectious Disease Research Collaboration (IDRC) PlotMbale Clinical Research Institute (MCRI)Faculty of Natural Resources and Environmental Sciences, Busitema UniversityFaculty of Engineering, Main Campus, Busitema UniversityMoroto Regional Referral HospitalSoroti Regional Referral HospitalMbale Clinical Research Institute (MCRI)Apac HospitalApac HospitalMbale Clinical Research Institute (MCRI)Mbale Clinical Research Institute (MCRI)Mbale Clinical Research Institute (MCRI)Infectious Disease Research Collaboration (IDRC) PlotUganda Ministry of Health, Central Public Health LaboratoryUganda Ministry of Health, Central Public Health LaboratoryInfectious Disease Research Collaboration (IDRC) PlotAbstract Background In Uganda, antimalarial resistance in Plasmodium falciparum poses serious public health and treatment challenges. Globally, recent data have highlighted the roles of following genes in malaria resistance: Plasmodium falciparum dihydrofolate reductase (Pfdhfr), Plasmodium falciparum dihydropteroate synthetase (Pfdhps), Plasmodium falciparum chloroquine resistance transporter (Pfcrt), Plasmodium falciparum multidrug resistance gene 1 (Pfmdr1), and Plasmodium falciparum K13 propeller domain (Pfk13). This study investigated the prevalence and characteristics of P. falciparum molecular markers linked to antimalarial resistance in Northern, Northeastern, and Eastern Uganda. Methods This cross-sectional study collected 200 dried blood samples from children (2 months to 12 years) in Northern, Eastern, and Northeastern Uganda. Samples were from malaria-positive cases confirmed by rapid diagnostic tests and microscopy. Genomic DNA was extracted from these samples and analysed using Molecular Inversion Probes to detect Plasmodium falciparum genetic mutations. The sequencing was performed on the Illumina MiSeq platform, and raw data was organized and analysed with MIPTools software. Results The study sequenced over 50% of the samples at each site as follows: Apac 87.7% (43/49), Moroto 68.0% (34/50), Soroti 65.0% (13/20) and Mbale 53.1% (43/81). The Pfk13 A675V and C469Y mutations varied from 0 to 23.3% and 8.3–14.3%, in four sites, with consistently low prevalence in Apac. The Pfdhfr N51I and S108N mutations were fixed in all districts, while C59R was fixed in Moroto and nearing fixation (92–97%) in other regions. The emerging I164L mutation ranged from 1 to 10% in all sites. The Pfdhps A437G and K540E mutations were fixed in Soroti, with 3–5% wild-type prevalence in other sites. The A581G mutation showed 2.3% mixed genotypes in Mbale only. The Pfcrt K76T was predominantly wild type, except for 5% mutants in Mbale and Moroto. The pfmdr1 N86Y were wild type across all districts, except for 15% mixed genotypes in Soroti. Conclusion This study reveal rising partial artemisinin resistance and widespread antifolate resistance surpassing WHO thresholds in Northern, Northeastern, and Eastern Uganda. Emerging super-resistant parasites pose a serious threat to malaria control, necessitating urgent enhanced surveillance and alternative treatment strategies.https://doi.org/10.1186/s12936-025-05439-xAntimalarial resistanceUgandaPfk13PfdhfrPfdhps Pfmdr1Pfcrt
spellingShingle Peter Olupot-Olupot
George Paasi
Thomas Katairo
Jimmy Patrick Alunyo
Alice Nakiyemba
Gilbert Gilibrays Ocen
Stephen Pande
Florance Alaroker
William Okiror
Emmaluel Ocen
Alex Oula
Charles Benard Okalebo
Ongodia Paul
Denis Amorut
Stephen Tukwasibwe
Susan Nabadda Ndidde
Isaac Sewanyana
Samuel L. Nsobya
Comprehensive analysis of molecular markers linked to antimalarial drug resistance in Plasmodium falciparum in Northern, Northeastern and Eastern Uganda
Malaria Journal
Antimalarial resistance
Uganda
Pfk13
Pfdhfr
Pfdhps Pfmdr1
Pfcrt
title Comprehensive analysis of molecular markers linked to antimalarial drug resistance in Plasmodium falciparum in Northern, Northeastern and Eastern Uganda
title_full Comprehensive analysis of molecular markers linked to antimalarial drug resistance in Plasmodium falciparum in Northern, Northeastern and Eastern Uganda
title_fullStr Comprehensive analysis of molecular markers linked to antimalarial drug resistance in Plasmodium falciparum in Northern, Northeastern and Eastern Uganda
title_full_unstemmed Comprehensive analysis of molecular markers linked to antimalarial drug resistance in Plasmodium falciparum in Northern, Northeastern and Eastern Uganda
title_short Comprehensive analysis of molecular markers linked to antimalarial drug resistance in Plasmodium falciparum in Northern, Northeastern and Eastern Uganda
title_sort comprehensive analysis of molecular markers linked to antimalarial drug resistance in plasmodium falciparum in northern northeastern and eastern uganda
topic Antimalarial resistance
Uganda
Pfk13
Pfdhfr
Pfdhps Pfmdr1
Pfcrt
url https://doi.org/10.1186/s12936-025-05439-x
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