title: Absence of in vivo selection for K13 mutations after artemether-lumefantrine treatment in Uganda.
abstract: Individual drug treatment may select resistant parasites in the human body, a process termed in vivo selection. Some single nucleotide polymorphisms in Plasmodium falciparum chloroquine-resistance transporter (pfcrt) and multidrug resistance gene 1 (pfmdr1) genes have been reportedly selected after artemether-lumefantrine treatment. However, there is a paucity of data regarding in vivo selection of P. falciparum Kelch propeller domain (pfkelch13) polymorphisms, responsible for artemisinin-resistance in Asia, and six putative background mutations for artemisinin resistance; D193Y in ferredoxin, T484I in multiple resistance protein 2, V127M in apicoplast ribosomal protein S10, I356T in pfcrt, V1157L in protein phosphatase and C1484F in phosphoinositide-binding protein.
  
  
  
  Artemether-lumefantrine efficacy study with a follow-up period of 28 days was conducted in northern Uganda in 2014. The above-mentioned genotypes were comparatively analysed before drug administration and on days; 3, 7, and 28 days after treatment.
  
  
  
  In 61 individuals with successful follow-up, artemether-lumefantrine treatment regimen was very effective with PCR adjusted efficacy of 95.2%. Among 146 isolates obtained before treatment, wild-type alleles were observed in 98.6% of isolates in pfkelch13 and in all isolates in the six putative background genes except I356T in pfcrt, which had 2.4% of isolates as mixed infections. In vivo selection study revealed that all isolates detected in the follow-up period harboured wild type alleles in pfkelch13 and the six background genes.
  
  
  
  Mutations in pfkelch13 and the six background genes may not play an important role in the in vivo selection after artemether-lumefantrine treatment in Uganda. Different mechanisms might rather be associated with the existence of parasites after treatment.
authors.affiliation: Division of Malaria Research, Proteo-Science Center, Ehime University, 3 Bunkyo-cho, Matsuyama, Ehime, 790-8577, Japan.
authors.forename: Betty
authors.initials: B
authors.lastname: Balikagala
authors.affiliation: Department of Molecular and Cellular Parasitology, School of Medicine, Juntendo University, Tokyo, 113-8421, Japan. tmita@juntendo.ac.jp.
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Mita
authors.affiliation: Department of Molecular and Cellular Parasitology, School of Medicine, Juntendo University, Tokyo, 113-8421, Japan.
authors.forename: Mie
authors.initials: M
authors.lastname: Ikeda
authors.affiliation: Department of International Affairs and Tropical Medicine, School of Medicine, Tokyo Women's Medical University, Tokyo, Japan.
authors.forename: Miki
authors.initials: M
authors.lastname: Sakurai
authors.affiliation: Health Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Takamatsu, Japan.
authors.forename: Shouki
authors.initials: S
authors.lastname: Yatsushiro
authors.affiliation: Department of International Affairs and Tropical Medicine, School of Medicine, Tokyo Women's Medical University, Tokyo, Japan.
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Takahashi
authors.affiliation: Department of Molecular and Cellular Parasitology, School of Medicine, Juntendo University, Tokyo, 113-8421, Japan.
authors.forename: Shin-Ichiro
authors.initials: SI
authors.lastname: Tachibana
authors.affiliation: St. Mary's Hospital LACOR, Gulu, Uganda.
authors.forename: Mary
authors.initials: M
authors.lastname: Auma
authors.affiliation: Division of Malaria Research, Proteo-Science Center, Ehime University, 3 Bunkyo-cho, Matsuyama, Ehime, 790-8577, Japan.
authors.forename: Edward H
authors.initials: EH
authors.lastname: Ntege
authors.affiliation: Division of Malaria Research, Proteo-Science Center, Ehime University, 3 Bunkyo-cho, Matsuyama, Ehime, 790-8577, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Ito
authors.affiliation: Division of Malaria Research, Proteo-Science Center, Ehime University, 3 Bunkyo-cho, Matsuyama, Ehime, 790-8577, Japan.
authors.forename: Eizo
authors.initials: E
authors.lastname: Takashima
authors.affiliation: Department of Molecular Protozoology, Research Institute for Microbial Diseases, Osaka University, Suita, Japan.
authors.forename: Nirianne Marie Q
authors.initials: NM
authors.lastname: Palacpac
authors.affiliation: Med Biotech Laboratories, Kampala, Uganda.
authors.forename: Thomas G
authors.initials: TG
authors.lastname: Egwang
authors.affiliation: Department of Biology, Faculty of Science, Gulu University, Gulu, Uganda.
authors.forename: Joseph Okello
authors.initials: JO
authors.lastname: Onen
authors.affiliation: Health Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Takamatsu, Japan.
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Kataoka
authors.affiliation: Department of Molecular Protozoology, Research Institute for Microbial Diseases, Osaka University, Suita, Japan.
authors.forename: Eisaku
authors.initials: E
authors.lastname: Kimura
authors.affiliation: Department of Molecular Protozoology, Research Institute for Microbial Diseases, Osaka University, Suita, Japan.
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Horii
authors.affiliation: Division of Malaria Research, Proteo-Science Center, Ehime University, 3 Bunkyo-cho, Matsuyama, Ehime, 790-8577, Japan. tsuboi.takafumi.mb@ehime-u.ac.jp.
authors.forename: Takafumi
authors.initials: T
authors.lastname: Tsuboi
mesh_terms: D000293:Adolescent; D000328:Adult; D000962:Antimalarials / Q000627:therapeutic use*; D000077611:Artemether, Lumefantrine Drug Combination; D037621:Artemisinins / Q000627:therapeutic use*; D002648:Child; D002675:Child, Preschool; D004338:Drug Combinations; D004351:Drug Resistance; D004983:Ethanolamines / Q000627:therapeutic use*; D005260:Female; D005449:Fluorenes / Q000627:therapeutic use*; D006801:Humans; D007223:Infant; D016778:Malaria, Falciparum / Q000188:drug therapy* / Q000469:parasitology; D008297:Male; D009154:Mutation; D010963:Plasmodium falciparum / Q000187:drug effects* / Q000302:isolation & purification; D011110:Polymorphism, Genetic; D015800:Protozoan Proteins / Q000235:genetics*; D012641:Selection, Genetic; D014454:Uganda; D055815:Young Adult
chemical_list: D000962:Antimalarials; D000077611:Artemether, Lumefantrine Drug Combination; D037621:Artemisinins; D004338:Drug Combinations; D004983:Ethanolamines; D005449:Fluorenes; D015800:Protozoan Proteins
keywords: Artemether–lumefantrine; Drug resistance; In vivo selection; Plasmodium falciparum; Polymorphisms; pfkelch13