{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,8]],"date-time":"2026-05-08T05:55:16Z","timestamp":1778219716830,"version":"3.51.4"},"reference-count":33,"publisher":"Public Library of Science (PLoS)","issue":"5","license":[{"start":{"date-parts":[[2022,5,5]],"date-time":"2022-05-05T00:00:00Z","timestamp":1651708800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>In Uganda, Artemether-Lumefantrine and Artesunate are recommended for uncomplicated and severe malaria respectively, but are currently threatened by parasite resistance. Genetic and epigenetic factors play a role in predisposing <jats:italic>Plasmodium falciparum<\/jats:italic> parasites to acquiring <jats:italic>Pfkelch13<\/jats:italic> (K13) mutations associated with delayed artemisinin parasite clearance as reported in Southeast Asia. In this study, we report on the prevalence of mutations in the K13, <jats:italic>pfmdr-2<\/jats:italic> (<jats:italic>P<\/jats:italic>. <jats:italic>falciparum<\/jats:italic> multidrug resistance protein 2), <jats:italic>fd<\/jats:italic> (ferredoxin), <jats:italic>pfcrt<\/jats:italic> (<jats:italic>P<\/jats:italic>. <jats:italic>falciparum<\/jats:italic> chloroquine resistance transporter), and <jats:italic>arps10<\/jats:italic> (apicoplast ribosomal protein S10) genes in <jats:italic>Plasmodium falciparum<\/jats:italic> parasites prior to (2005) and after (2013) introduction of artemisinin combination therapies for malaria treatment in Uganda. A total of 200 <jats:italic>P<\/jats:italic>. <jats:italic>falciparum<\/jats:italic> parasite DNA samples were screened. Parasite DNA was extracted using QIAamp DNA mini kit (Qiagen, GmbH, Germany) procedure. The PCR products were sequenced using Sanger dideoxy sequencing method. Of the 200 <jats:italic>P<\/jats:italic>. <jats:italic>falciparum<\/jats:italic> DNA samples screened, sequencing for mutations in <jats:italic>K13<\/jats:italic>, <jats:italic>pfmdr-2<\/jats:italic>, <jats:italic>fd<\/jats:italic>, <jats:italic>pfcrt<\/jats:italic>, <jats:italic>arps10<\/jats:italic> genes was successful in 142, 186, 141, 128 and 74 samples respectively. Overall, we detected six (4.2%, 6\/142; 95%CI: 1.4\u20137.0) K13 single nucleotide polymorphisms (SNPs), of which 3.9% (2\/51), 4.4% (4\/91) occurred in 2005 and 2013 samples respectively. All four K13 SNPs in 2013 samples were non-synonymous (A578S, E596V, S600C and E643K) while of the two SNPs in 2005 samples, one (Y588N) is non-synonymous and the other (I587I) is synonymous. There was no statistically significant difference in the prevalence of K13 (<jats:italic>p<\/jats:italic> = 0.112) SNPs in the samples collected in 2005 and 2013. The overall prevalence of SNPs in <jats:italic>pfmdr-2<\/jats:italic> gene was 39.8% (74\/186, 95%CI: 25.1\u201350.4). Of this, 4.2% (4\/95), 76.9% (70\/91) occurred in 2005 and 2013 samples respectively. In 2005 samples only one SNP, Y423F (4.2%, 4\/95) was found while in 2013, Y423F (38.5%, 35\/91) and I492V (38.5%, 35\/91) SNPs in the <jats:italic>pfmdr-2<\/jats:italic> gene were found. There was a statistically significant difference in the prevalence of <jats:italic>pfmdr-2<\/jats:italic> SNPs in the samples collected in 2005 and 2013 (<jats:italic>p<\/jats:italic>&lt;0.001). The overall prevalence of <jats:italic>arps10<\/jats:italic> mutations was 2.7% (2\/72, 95%CI: 0.3\u20134.2). Two mutations, V127M (4.5%: 1\/22) and D128H (4.5%: 1\/22) in the <jats:italic>arps10<\/jats:italic> gene were each found in <jats:italic>P<\/jats:italic>. <jats:italic>falciparum<\/jats:italic> parasite samples collected in 2013. There was no statistically significant difference in the prevalence of <jats:italic>arps10<\/jats:italic> SNPs in the samples collected in 2005 and 2013 (<jats:italic>p<\/jats:italic> = 0.238). There were more <jats:italic>pfmdr-2<\/jats:italic> SNPs in <jats:italic>P<\/jats:italic>. <jats:italic>falciparum<\/jats:italic> parasites collected after introduction of Artemisinin combination therapies in malaria treatment. This is an indicator of the need for continuous surveillance to monitor emergence of molecular markers of artemisinin resistance and its potential drivers in malaria affected regions globally.<\/jats:p>","DOI":"10.1371\/journal.pone.0268095","type":"journal-article","created":{"date-parts":[[2022,5,5]],"date-time":"2022-05-05T17:39:05Z","timestamp":1651772345000},"page":"e0268095","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":13,"title":["Prevalence of arps10, fd, pfmdr-2, pfcrt and pfkelch13 gene mutations in Plasmodium falciparum parasite population in Uganda"],"prefix":"10.1371","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8223-5465","authenticated-orcid":true,"given":"Moses","family":"Ocan","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fred Katabazi","family":"Ashaba","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Savannah","family":"Mwesigwa","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kigozi","family":"Edgar","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Moses R.","family":"Kamya","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sam L.","family":"Nsobya","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2022,5,5]]},"reference":[{"key":"pone.0268095.ref001","volume-title":"World Health Organization: 20 years of global progress and challenges","author":"World Health Organization","year":"2020"},{"issue":"3","key":"pone.0268095.ref002","doi-asserted-by":"crossref","first-page":"753","DOI":"10.4269\/ajtmh.17-0345","article-title":"Plasmodium Species Infecting Children Presenting with Malaria in Uganda","volume":"97","author":"V Asua","year":"2017","journal-title":"Am J Trop Med Hyg"},{"key":"pone.0268095.ref003","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1186\/s12936-017-1680-8","article-title":"Rapid decline in the susceptibility of Plasmodium falciparum to dihydroartemisinin\u2013piperaquine in the south of Vietnam","volume":"16","author":"NV Thanh","year":"2017","journal-title":"Malaria Journal"},{"issue":"2","key":"pone.0268095.ref004","doi-asserted-by":"crossref","first-page":"818","DOI":"10.1128\/AAC.00686-12","article-title":"Efficacy of dihydroartemisinin-piperaquine for treatment of uncomplicated Plasmodium falciparum and Plasmodium vivax in Cambodia, 2008 to 2010","volume":"57","author":"R Leang","year":"2013","journal-title":"Antimicrob Agents Chemother"},{"key":"pone.0268095.ref005","unstructured":"Ministry of Health. UGANDA NATIONAL MALARIA CONTROL POLICY. In: National Malaria Control Program Moh, editor. Kampala: Ministry of Health; 2011. p. 1\u201319."},{"key":"pone.0268095.ref006","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1186\/s12936-021-03711-4","article-title":"Efcacy and safety of dihydroartemisinin\u2013piperaquine versus artemether\u2013lumefantrine for treatment of uncomplicated Plasmodium falciparum malaria in Ugandan children: a systematic review and meta-analysis of randomized control trials","volume":"20","author":"DG Assefa","year":"2021","journal-title":"Malar J"},{"key":"pone.0268095.ref007","doi-asserted-by":"crossref","first-page":"1326","DOI":"10.1086\/651562","article-title":"High heritability of malaria parasite clearance rate indicates a genetic basis for artemisinin resistance in Western Cambodia","volume":"201","author":"TJ Anderson","year":"2010","journal-title":"Journal of Infectious Diseases"},{"issue":"3","key":"pone.0268095.ref008","doi-asserted-by":"crossref","first-page":"226","DOI":"10.1038\/ng.3189","article-title":"Genetic architecture of artemisinin-resistant Plasmodium falciparum","volume":"47","author":"O Miotto","year":"2015","journal-title":"Nat Genet"},{"issue":"5","key":"pone.0268095.ref009","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1056\/NEJMoa0808859","article-title":"Artemisinin resistance in Plasmodium falciparum malaria","volume":"361","author":"AM Dondorp","year":"2009","journal-title":"N Engl J Med"},{"key":"pone.0268095.ref010","doi-asserted-by":"crossref","first-page":"2453","DOI":"10.1056\/NEJMoa1513137","article-title":"A Worldwide Map of Plasmodium falciparum K13-Propeller Polymorphisms","volume":"374","author":"D M\u00e9nard","year":"2016","journal-title":"N Engl J Med"},{"issue":"5","key":"pone.0268095.ref011","doi-asserted-by":"crossref","first-page":"670","DOI":"10.1093\/infdis\/jiu491","article-title":"Independent emergence of artemisinin resistance mutations among Plasmodium falciparum in Southeast Asia","volume":"211","author":"S Takala-Harrison","year":"2015","journal-title":"J Infect Dis"},{"key":"pone.0268095.ref012","doi-asserted-by":"crossref","first-page":"472","DOI":"10.1186\/1475-2875-13-472","article-title":"Limited polymorphisms in k13 gene in Plasmodium falciparum isolates from Dakar, Senegal in 2012\u20132013","volume":"13","author":"M Torrentino-Madamet","year":"2014","journal-title":"Malaria Journal"},{"key":"pone.0268095.ref013","doi-asserted-by":"crossref","first-page":"452","DOI":"10.1186\/1475-2875-13-452","article-title":"Artemisinin resistance\u2014modelling the potential human and economic costs","volume":"13","author":"Y Lubell","year":"2014","journal-title":"Malaria Journal"},{"key":"pone.0268095.ref014","first-page":"689","article-title":"Impact of chloroquine resistance on malaria mortality","volume":"321","author":"JF Trape","year":"1998","journal-title":"C R Acad Sci Paris Series III"},{"issue":"9","key":"pone.0268095.ref015","doi-asserted-by":"crossref","first-page":"682","DOI":"10.1016\/j.pt.2016.05.010","article-title":"Artemisinin Action and Resistance in Plasmodium falciparum","volume":"32","author":"L Tilley","year":"2016","journal-title":"Trends Parasitol"},{"key":"pone.0268095.ref016","article-title":"Changing Prevalence of Potential Mediators of Aminoquinoline, Antifolate, and Artemisinin Resistance Across Uganda","author":"V Asua","year":"2020","journal-title":"The Journal of Infectious Diseases"},{"issue":"1","key":"pone.0268095.ref017","doi-asserted-by":"crossref","first-page":"294","DOI":"10.3201\/eid2701.203527","article-title":"Increase in Kelch 13 Polymorphisms in Plasmodium falciparum, Southern Rwanda","volume":"27","author":"C Bergmann","year":"2021","journal-title":"Emerging Infectious Diseases"},{"key":"pone.0268095.ref018","doi-asserted-by":"crossref","first-page":"1","DOI":"10.4061\/2011\/683167","article-title":"Malaria Treatment Policy Change and Implementation: The Case of Uganda","volume":"2011","author":"M Nanyunja","year":"2011","journal-title":"Malaria Research and Treatment"},{"issue":"20","key":"pone.0268095.ref019","doi-asserted-by":"crossref","first-page":"2210","DOI":"10.1001\/jama.297.20.2210","article-title":"Combination Therapy for Uncomplicated Falciparum Malaria in Ugandan Children A Randomized Trial","volume":"297","author":"G Dorsey","year":"2007","journal-title":"JAMA"},{"issue":"5","key":"pone.0268095.ref020","doi-asserted-by":"crossref","first-page":"903","DOI":"10.4269\/ajtmh.14-0312","article-title":"Malaria Transmission, Infection, and Disease at Three Sites with Varied Transmission Intensity in Uganda: Implications for Malaria Control","volume":"92","author":"MR Kamya","year":"2015","journal-title":"Am J Trop Med Hyg"},{"key":"pone.0268095.ref021","volume-title":"Tutor\u2019s guide","author":"World Health Organization","year":"2004","edition":"2"},{"key":"pone.0268095.ref022","doi-asserted-by":"crossref","first-page":"564","DOI":"10.1016\/S0140-6736(94)91520-2","article-title":"Diagnosis of malaria by detection of Plasmodium falciparum HRP-2 antigen with a rapid dipstick antigen-capture assay","volume":"343","author":"C Beadle","year":"1994","journal-title":"Lancet"},{"issue":"7481","key":"pone.0268095.ref023","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1038\/nature12876","article-title":"A molecular marker of artemisinin-resistant Plasmodium falciparum malaria","volume":"505","author":"F Ariey","year":"2014","journal-title":"Natue"},{"issue":"1","key":"pone.0268095.ref024","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s12936-021-03613-5","article-title":"Molecular surveillance of pfcrt, pfmdr1 and pfk13-propeller mutations in Plasmodium falciparum isolates imported from Africa to China","volume":"20","author":"F Huang","year":"2021","journal-title":"Malar J"},{"issue":"2","key":"pone.0268095.ref025","doi-asserted-by":"crossref","first-page":"453","DOI":"10.5114\/aoms.2019.84825","article-title":"Identification of seven novel variants in the \u03b2-globin gene in transfusion-dependent and normal patients","volume":"16","author":"SA Aldakeel","year":"2020","journal-title":"Archives of Medical Science"},{"key":"pone.0268095.ref026","doi-asserted-by":"crossref","first-page":"428","DOI":"10.1126\/science.1260867","article-title":"Drug resistance. K13-propeller mutations confer artemisinin resistance in Plasmodium falciparum clinical isolates","volume":"347","author":"J Straimer","year":"2015","journal-title":"Science"},{"issue":"12","key":"pone.0268095.ref027","doi-asserted-by":"crossref","first-page":"e1009266","DOI":"10.1371\/journal.pgen.1009266","article-title":"Genetic background and PfKelch13 affect artemisinin susceptibility of PfCoronin mutants in Plasmodium falciparum","volume":"16","author":"AI Sharma","year":"2020","journal-title":"PLoS Genet"},{"issue":"1","key":"pone.0268095.ref028","doi-asserted-by":"crossref","first-page":"e0245336","DOI":"10.1371\/journal.pone.0245336","article-title":"Antimalarial drug resistance molecular markers of Plasmodium falciparum isolates from Sudan during 2015\u20132017","volume":"16","author":"M Hussien","year":"2021","journal-title":"PloS ONE"},{"issue":"8","key":"pone.0268095.ref029","doi-asserted-by":"crossref","first-page":"e0235401","DOI":"10.1371\/journal.pone.0235401","article-title":"Antimalarial drug resistance molecular makers of Plasmodium falciparum isolates from Sudan during 2015\u20132017","volume":"15","author":"M Hussien","year":"2020","journal-title":"PLoS ONE"},{"key":"pone.0268095.ref030","doi-asserted-by":"crossref","first-page":"e01234","DOI":"10.1128\/AAC.01234-19","article-title":"The Diversity of the Plasmodium falciparum K13 Propeller Domain Did Not Increase after Implementation of ArtemisininBased Combination Therapy in Uganda","volume":"63","author":"MD Conrad","year":"2019","journal-title":"Antimicrob Agents Chemother"},{"issue":"10","key":"pone.0268095.ref031","doi-asserted-by":"crossref","first-page":"1602","DOI":"10.1038\/s41591-020-1005-2","article-title":"Emergence and clonal expansion of in vitro artemisinin-resistant Plasmodium falciparum kelch13 R561H mutant parasites in Rwanda","volume":"26","author":"A Uwimana","year":"2020","journal-title":"Nat Med"},{"issue":"4","key":"pone.0268095.ref032","doi-asserted-by":"crossref","first-page":"718","DOI":"10.3201\/eid2404.170141","article-title":"Artemisinin-Resistant Plasmodium falciparum with High Survival Rates, Uganda, 2014\u20132016","volume":"24","author":"M Ikeda","year":"2018","journal-title":"Emerg Infect Dis"},{"key":"pone.0268095.ref033","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1056\/NEJMoa2101746","article-title":"Evidence of Artemisinin-Resistant Malaria in Africa","volume":"385","author":"B Balikagala","year":"2021","journal-title":"N ENGL J MED"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0268095","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,5,5]],"date-time":"2022-05-05T17:39:51Z","timestamp":1651772391000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0268095"}},"subtitle":[],"editor":[{"given":"Luzia Helena","family":"Carvalho","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2022,5,5]]},"references-count":33,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2022,5,5]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0268095","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,5,5]]}}}