Detection of atovaquone and MalaroneTM resistance conferring mutations in Plasmodium falciparum cytochrome b gene (cytb)

J. P. Gil, F Nogueira, J. Strömberg-Nörklit, J. Lindberg, M. Carrolo, C. Casimiro, D Lopes, Ana Paula Arez, Pedro Vitor Lemos Cravo, Virgilio Estólio do Rosário

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42 Citations (Scopus)

Abstract

Clinical treatment failures of the hydroxynaphthoquinone atovaquone or its combination with proguanil (Malarone) in Plasmodium falciparum malaria has been recently documented. These events have been associated to single nucleotide polymorphisms (SNPs) in the parasite cytochrome b gene (cytb). In this report we describe a set of nest PCR-RFLP methods developed for the fast detection of all known cytb mutations associated to resistance to these drugs. The methods were successfully applied for the analysis of phenol-chloroform extracted DNA samples from patients not cured by Malarone, and from an established parasite clone. Further, the protocol for the detection of the A803C mutation was applied to 164 DNA field samples extracted through crude methanol-based protocols, originated from several malaria settings. The PCR-RFLP methods here presented can be used as a valuable for the clinical detection and study of Malarone and atovaquone P. falciparum resistance.
Original languageEnglish
Pages (from-to)85-89
Number of pages5
JournalMolecular and Cellular Probes
Volume17
Issue number2-3
DOIs
Publication statusPublished - Apr 2003

Keywords

  • Plasmodium falciparum
  • Resistance
  • Cytochrome b
  • Atovaquone
  • Malarone
  • PCR-RFLP

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