Prevalence of Plasmodium falciparum parasites harbouring chloroquine-resistant but not artemisinin-resistant alleles in Busia County, Western Kenya

Abstract Background The recent emergence of Plasmodium falciparum parasites harbouring kelch 13 mutations associated with artemisinin partial resistance (ART-R) in sub-Saharan Africa is a major concern. Regular molecular epidemiological surveys are recommended to monitor the level of mutant parasite...

Full description

Saved in:
Bibliographic Details
Main Authors: Noah Machuki Onchieku, Nemrod Gesusu, Emmanuelle Caspar, Lewis Karani, Kelvin Thiong’o, Luna Kamau, Daniel Kiboi, Lucas Thiebaut, Laurence Ma, Francis Kimani, Damaris Matoke-Muhia, Didier Ménard
Format: Article
Language:English
Published: BMC 2025-07-01
Series:Malaria Journal
Subjects:
Online Access:https://doi.org/10.1186/s12936-025-05486-4
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849333691026767872
author Noah Machuki Onchieku
Nemrod Gesusu
Emmanuelle Caspar
Lewis Karani
Kelvin Thiong’o
Luna Kamau
Daniel Kiboi
Lucas Thiebaut
Laurence Ma
Francis Kimani
Damaris Matoke-Muhia
Didier Ménard
author_facet Noah Machuki Onchieku
Nemrod Gesusu
Emmanuelle Caspar
Lewis Karani
Kelvin Thiong’o
Luna Kamau
Daniel Kiboi
Lucas Thiebaut
Laurence Ma
Francis Kimani
Damaris Matoke-Muhia
Didier Ménard
author_sort Noah Machuki Onchieku
collection DOAJ
description Abstract Background The recent emergence of Plasmodium falciparum parasites harbouring kelch 13 mutations associated with artemisinin partial resistance (ART-R) in sub-Saharan Africa is a major concern. Regular molecular epidemiological surveys are recommended to monitor the level of mutant parasites associated with drug resistance. Objective To analyse single nucleotide polymorphisms (SNPs) in P. falciparum genes associated with resistance to antimalarial drugs in blood samples collected from malaria patients prior to treatment in Busia County, Western Kenya, using a highly multiplexed deep amplicon sequencing protocol. Methods Parasite DNA was extracted from dried blood spots (DBS) collected from patients attending health facilities along the Kenya-Uganda border, in Busia County. A highly multiplexed deep amplicon sequencing protocol was applied to analyze SNPs in genes associated with antimalarial drugs resistance. Results Out of 118 analysed samples, Pfkelch13 mutations were identified in 6 (5.1%) samples. All the six samples harboured A578S, one of the most frequently detected Pfkelch13 mutations in Africa, which does not confer ART-R. The data also indicated that 11% (13) of the samples had mutant Pfcrt alleles, with 62% harbouring the triple mutant haplotype, CVIET. In contrast, 98% of the samples had dhfr mutations at positions N51I, C59R, or S108N alleles. All the samples carried mutations in the dhps gene with a high prevalence of the single A437G mutation (84%) and a lower frequency of double or triple mutations. Further analysis revealed absence of deletions in the Pfhrp2, with no mutations at Pfmdr1 codons 86 or 1246, both of which are associated with chloroquine resistance. Conclusion These findings show absence of validated Pfkelch13 markers associated with ART-R in Busia County, Western Kenya. Although chloroquine-sensitive (wild-type) parasites are widely circulating, sulfadoxine-pyrimethamine-sensitive parasites remain rare. Continued large scale genomic surveillance studies are essential to detect emerging resistance and guide malaria treatment strategies in the region.
format Article
id doaj-art-5d4000dcc8e04bd2b2ca2d7a8c1797ad
institution Kabale University
issn 1475-2875
language English
publishDate 2025-07-01
publisher BMC
record_format Article
series Malaria Journal
spelling doaj-art-5d4000dcc8e04bd2b2ca2d7a8c1797ad2025-08-20T03:45:47ZengBMCMalaria Journal1475-28752025-07-0124111110.1186/s12936-025-05486-4Prevalence of Plasmodium falciparum parasites harbouring chloroquine-resistant but not artemisinin-resistant alleles in Busia County, Western KenyaNoah Machuki Onchieku0Nemrod Gesusu1Emmanuelle Caspar2Lewis Karani3Kelvin Thiong’o4Luna Kamau5Daniel Kiboi6Lucas Thiebaut7Laurence Ma8Francis Kimani9Damaris Matoke-Muhia10Didier Ménard11Sports Science Research Centre, Kenya Medical Research Institute (KEMRI)Centre for Biotechnology Research Development (CBRD), Kenya Medical Research Institute (KEMRI)Malaria Genetics and Resistance Team (MEGATEAM), UR 3073-Pathogens Host Arthropods Vectors Interactions Unit, Université de StrasbourgCentre for Biotechnology Research Development (CBRD), Kenya Medical Research Institute (KEMRI)Centre for Biotechnology Research Development (CBRD), Kenya Medical Research Institute (KEMRI)Centre for Biotechnology Research Development (CBRD), Kenya Medical Research Institute (KEMRI)Department of Biochemistry, Jomo Kenyatta University of Agriculture and Technology (JKUAT)Malaria Genetics and Resistance Team (MEGATEAM), UR 3073-Pathogens Host Arthropods Vectors Interactions Unit, Université de StrasbourgBiomics Platform, Institut Pasteur, Université Paris CiteCentre for Biotechnology Research Development (CBRD), Kenya Medical Research Institute (KEMRI)Centre for Biotechnology Research Development (CBRD), Kenya Medical Research Institute (KEMRI)Malaria Genetics and Resistance Team (MEGATEAM), UR 3073-Pathogens Host Arthropods Vectors Interactions Unit, Université de StrasbourgAbstract Background The recent emergence of Plasmodium falciparum parasites harbouring kelch 13 mutations associated with artemisinin partial resistance (ART-R) in sub-Saharan Africa is a major concern. Regular molecular epidemiological surveys are recommended to monitor the level of mutant parasites associated with drug resistance. Objective To analyse single nucleotide polymorphisms (SNPs) in P. falciparum genes associated with resistance to antimalarial drugs in blood samples collected from malaria patients prior to treatment in Busia County, Western Kenya, using a highly multiplexed deep amplicon sequencing protocol. Methods Parasite DNA was extracted from dried blood spots (DBS) collected from patients attending health facilities along the Kenya-Uganda border, in Busia County. A highly multiplexed deep amplicon sequencing protocol was applied to analyze SNPs in genes associated with antimalarial drugs resistance. Results Out of 118 analysed samples, Pfkelch13 mutations were identified in 6 (5.1%) samples. All the six samples harboured A578S, one of the most frequently detected Pfkelch13 mutations in Africa, which does not confer ART-R. The data also indicated that 11% (13) of the samples had mutant Pfcrt alleles, with 62% harbouring the triple mutant haplotype, CVIET. In contrast, 98% of the samples had dhfr mutations at positions N51I, C59R, or S108N alleles. All the samples carried mutations in the dhps gene with a high prevalence of the single A437G mutation (84%) and a lower frequency of double or triple mutations. Further analysis revealed absence of deletions in the Pfhrp2, with no mutations at Pfmdr1 codons 86 or 1246, both of which are associated with chloroquine resistance. Conclusion These findings show absence of validated Pfkelch13 markers associated with ART-R in Busia County, Western Kenya. Although chloroquine-sensitive (wild-type) parasites are widely circulating, sulfadoxine-pyrimethamine-sensitive parasites remain rare. Continued large scale genomic surveillance studies are essential to detect emerging resistance and guide malaria treatment strategies in the region.https://doi.org/10.1186/s12936-025-05486-4MalariaPlasmodium falciparumArtemisininChloroquineDrug resistanceMultiplexed deep amplicon sequencing
spellingShingle Noah Machuki Onchieku
Nemrod Gesusu
Emmanuelle Caspar
Lewis Karani
Kelvin Thiong’o
Luna Kamau
Daniel Kiboi
Lucas Thiebaut
Laurence Ma
Francis Kimani
Damaris Matoke-Muhia
Didier Ménard
Prevalence of Plasmodium falciparum parasites harbouring chloroquine-resistant but not artemisinin-resistant alleles in Busia County, Western Kenya
Malaria Journal
Malaria
Plasmodium falciparum
Artemisinin
Chloroquine
Drug resistance
Multiplexed deep amplicon sequencing
title Prevalence of Plasmodium falciparum parasites harbouring chloroquine-resistant but not artemisinin-resistant alleles in Busia County, Western Kenya
title_full Prevalence of Plasmodium falciparum parasites harbouring chloroquine-resistant but not artemisinin-resistant alleles in Busia County, Western Kenya
title_fullStr Prevalence of Plasmodium falciparum parasites harbouring chloroquine-resistant but not artemisinin-resistant alleles in Busia County, Western Kenya
title_full_unstemmed Prevalence of Plasmodium falciparum parasites harbouring chloroquine-resistant but not artemisinin-resistant alleles in Busia County, Western Kenya
title_short Prevalence of Plasmodium falciparum parasites harbouring chloroquine-resistant but not artemisinin-resistant alleles in Busia County, Western Kenya
title_sort prevalence of plasmodium falciparum parasites harbouring chloroquine resistant but not artemisinin resistant alleles in busia county western kenya
topic Malaria
Plasmodium falciparum
Artemisinin
Chloroquine
Drug resistance
Multiplexed deep amplicon sequencing
url https://doi.org/10.1186/s12936-025-05486-4
work_keys_str_mv AT noahmachukionchieku prevalenceofplasmodiumfalciparumparasitesharbouringchloroquineresistantbutnotartemisininresistantallelesinbusiacountywesternkenya
AT nemrodgesusu prevalenceofplasmodiumfalciparumparasitesharbouringchloroquineresistantbutnotartemisininresistantallelesinbusiacountywesternkenya
AT emmanuellecaspar prevalenceofplasmodiumfalciparumparasitesharbouringchloroquineresistantbutnotartemisininresistantallelesinbusiacountywesternkenya
AT lewiskarani prevalenceofplasmodiumfalciparumparasitesharbouringchloroquineresistantbutnotartemisininresistantallelesinbusiacountywesternkenya
AT kelvinthiongo prevalenceofplasmodiumfalciparumparasitesharbouringchloroquineresistantbutnotartemisininresistantallelesinbusiacountywesternkenya
AT lunakamau prevalenceofplasmodiumfalciparumparasitesharbouringchloroquineresistantbutnotartemisininresistantallelesinbusiacountywesternkenya
AT danielkiboi prevalenceofplasmodiumfalciparumparasitesharbouringchloroquineresistantbutnotartemisininresistantallelesinbusiacountywesternkenya
AT lucasthiebaut prevalenceofplasmodiumfalciparumparasitesharbouringchloroquineresistantbutnotartemisininresistantallelesinbusiacountywesternkenya
AT laurencema prevalenceofplasmodiumfalciparumparasitesharbouringchloroquineresistantbutnotartemisininresistantallelesinbusiacountywesternkenya
AT franciskimani prevalenceofplasmodiumfalciparumparasitesharbouringchloroquineresistantbutnotartemisininresistantallelesinbusiacountywesternkenya
AT damarismatokemuhia prevalenceofplasmodiumfalciparumparasitesharbouringchloroquineresistantbutnotartemisininresistantallelesinbusiacountywesternkenya
AT didiermenard prevalenceofplasmodiumfalciparumparasitesharbouringchloroquineresistantbutnotartemisininresistantallelesinbusiacountywesternkenya