title: Confirmation of emergence of mutations associated with atovaquone-proguanil resistance in unexposed Plasmodium falciparum isolates from Africa.
abstract: BACKGROUND
  In vitro and in vivo resistance of Plasmodium falciparum to atovaquone or atovaquone-proguanil hydrochloride combination has been associated to two point mutations in the parasite cytochrome b (cytb) gene (Tyr268Ser and Tyr268Asn). However, little is known about the prevalence of codon-268 mutations in natural populations of P. falciparum without previous exposure to the drug in Africa.
  
  
  METHODS
  The prevalence of codon-268 mutations in the cytb gene of African P. falciparum isolates from Nigeria, Malawi and Senegal, where atovaquone-proguanil has not been introduced for treatment of malaria was assessed. Genotyping of the cytb gene in isolates of P. falciparum was performed by PCR-restriction fragment length polymorphism and confirmed by sequencing.
  
  
  RESULTS
  295 samples from Nigeria (111), Malawi (91) and Senegal (93) were successfully analyzed for detection of either mutant Tyr268Ser or Tyr268Asn. No case of Ser268 or Asn268 was detected in cytb gene of parasites from Malawi or Senegal. However, Asn268 was detected in five out of 111 (4.5%) unexposed P. falciparum isolates from Nigeria. In addition, one out of these five mutant Asn268 isolates showed an additional cytb mutation leading to a Pro266Thr substitution inside the ubiquinone reduction site.
  
  
  CONCLUSIONS
  No Tyr268Ser mutation is found in cytb of P. falciparum isolates from Nigeria, Malawi or Senegal. This study reports for the first time cytb Tyr268Asn mutation in unexposed P. falciparum isolates from Nigeria. The emergence in Africa of P. falciparum isolates with cytb Tyr268Asn mutation is a matter of serious concern. Continuous monitoring of atovaquone-proguanil resistant P. falciparum in Africa is warranted for the rational use of this new antimalarial drug, especially in non-immune travelers.
authors.affiliation: Malaria Research Laboratories, Institute for Advanced Medical Research and Training, College of Medicine, University of Ibadan, Nigeria. chappi@hsph.harvard.edu
authors.forename: Christian T
authors.initials: CT
authors.lastname: Happi
authors.forename: Grace O
authors.initials: GO
authors.lastname: Gbotosho
authors.forename: Onikepe A
authors.initials: OA
authors.lastname: Folarin
authors.forename: Danny
authors.initials: D
authors.lastname: Milner
authors.forename: Ousmane
authors.initials: O
authors.lastname: Sarr
authors.forename: Akintunde
authors.initials: A
authors.lastname: Sowunmi
authors.forename: Dennis E
authors.initials: DE
authors.lastname: Kyle
authors.forename: Wilbur K
authors.initials: WK
authors.lastname: Milhous
authors.forename: Dyann F
authors.initials: DF
authors.lastname: Wirth
authors.forename: Ayoade M J
authors.initials: AM
authors.lastname: Oduola
mesh_terms: D000328:Adult; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D000962:Antimalarials / Q000494:pharmacology*; D053626:Atovaquone; D002648:Child; D003062:Codon / Q000235:genetics; D045303:Cytochromes b / Q000737:chemistry / Q000235:genetics*; D016054:DNA, Protozoan / Q000235:genetics; D004351:Drug Resistance / Q000235:genetics*; D006801:Humans; D016778:Malaria, Falciparum / Q000469:parasitology* / Q000517:prevention & control; D008295:Malawi; D008969:Molecular Sequence Data; D020125:Mutation, Missense; D009285:Naphthoquinones / Q000494:pharmacology*; D009549:Nigeria; D010963:Plasmodium falciparum / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D002727:Proguanil / Q000494:pharmacology*; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics*; D012675:Senegal; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014195:Travel
chemical_list: D000962:Antimalarials; D003062:Codon; D016054:DNA, Protozoan; D009285:Naphthoquinones; D015800:Protozoan Proteins; D045303:Cytochromes b; D002727:Proguanil; D053626:Atovaquone