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Multiple Phenotypic and Genotypic Artemisinin Sensitivity Evaluation of Malian Plasmodium falciparum Isolates
We assessed the ex vivo/in vitro sensitivity of 54 Malian Plasmodium falciparum isolates to artemisinin for the monitoring of drug resistance in this area. The artemisinin sensitivity of parasites was evaluated using 1) the ex vivo and in vitro parasite recrudescence detection after treatment of the ring stage with 1–200 nM artemisinin for 48 hours and 2) the in vitro parasite recrudescence kinetics assay over 7 days after 6-hour treatment of the ring stage with 700 nM dihydroartemisinin (DHA). In addition, as recommended by the World Health Organization for artemisinin resistance characterization, the ring-stage survival assay (RSA0–3 h) was performed and the parasite isolates were sequenced at the kelch 13 propeller locus. No clinical and molecular evidence of artemisinin resistance was observed. However, these isolates present different phenotypic profiles in response to artemisinin treatments. Despite all RSA0–3 h values less than 1.5%, six out of 46 (13.0%) isolates tested ex vivo and four out of six (66.7%) isolates tested in vitro were able to multiply after 48-hour treatments with 100 nM artemisinin. Moreover, five out of eight isolates tested showed faster parasite recovery after DHA treatment in kinetic assays. The presence of such phenotypes needs to be taken into account in the assessment of the efficacy of artemisinins in Mali. The assays presented here appear as valuable tools for the monitoring of artemisinin sensitivity in the field and thus could help to evaluate the risk of emergence of artemisinin resistance in Africa.
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Author Notes
These authors contributed equally to this work.
Financial support: This work was supported by the Fondation Mérieux and World Health Organization, Geneva; the French Centre National de la Recherche Scientifique; the French Institut National de la Santé et de la Recherche Médicale; the French Agence Nationale de la Recherche; and the ParaFrap Consortium.
Authors’ addresses: Karamoko Niaré and Ogobara K. Doumbo, Malaria Research and Training Center, University of Sciences, Techniques and Technologies of Bamako, Bamako, Mali, E-mails: karaniare@yahoo.fr and okd@icermali.org. Lucie Paloque, Pety Tor, Arba P. Ramadani, Jean-Michel Augereau, and Françoise Benoit-Vical, Laboratoire de Chimie de Coordination du CNRS, Centre Nationale de la Recherche Scientifique, Université de Toulouse, UPS, INPT, Toulouse, France, E-mails: lucie.paloque@lcc-toulouse.fr, smilyrab@yahoo.com, arba.pramundita@gmail.com, jean-michel.augereau@lcc-toulouse.fr, and francoise.vical@inserm.fr. Sandie Ménard, Centre de Physiopathologie de Toulouse Purpan, Université de Toulouse, Toulouse, France, E-mail: sandie.menard@inserm.fr. Antoine Dara, Malaria Research and Training Center, University of Sciences, Techniques and Technologies of Bamako, Bamako, Mali, and Division of Malaria Research, Institute for Global Health, University of Maryland School of Medicine, Baltimore, MD, E-mail: tonydara@icermali.org. Antoine Berry, Service de Parasitologie-Mycologie, Centre Hospitalier et Universitaire de Toulouse, Toulouse, France, and Centre de Physiopathologie de Toulouse Purpan, Université de Toulouse, Toulouse, France, E-mail: berry.a@chu-toulouse.fr.