Contributors: Institut Pasteur du Cambodge; Réseau International des Instituts Pasteur (RIIP); National Center for Parasitology, Entomology and Malaria Control Phnom Penh, Cambodia (CNM); Département Parasites et Insectes vecteurs - Department of Parasites and Insect Vectors; Institut Pasteur Paris (IP); WorldWide Antimalarial Resistance Network (WWARN); University of Oxford-Churchill Hospital Oxford Centre for Haematology; Centre for Tropical Medicine and Global Health Oxford, UK; Nuffield Department of Medicine Oxford, UK (Big Data Institute); University of Oxford-University of Oxford; This work was funded by grants from Institut Pasteur du Cambodge, USAID and BMGF through the World Health Organization, and Institut Pasteur Paris. PJG is member of the Worldwide Antimalarial Resistance Network supported by the Bill and Melinda Gates Foundation. VD is supported by a doctoral fellowship from the International Division, Institut Pasteur.; We thank all patients enrolled in the therapeutic efficacy studies, the health centres staff and the staff from the National Center for Parasitology, Entomology and Malaria Control in Cambodia for their support. We are grateful to Joel Tarning (Mahidol Oxford Tropical Medicine Research Unit, Bangkok, Thailand; Centre for Tropical Medicine, University of Oxford, Oxford, United Kingdom) who performed the piperaquine concentration measurements.
نبذة مختصرة : International audience ; BACKGROUND:The declining efficacy of dihydroartemisinin-piperaquine against Plasmodium falciparum in Cambodia, along with increasing numbers of recrudescent cases, suggests resistance to both artemisinin and piperaquine. Available in vitro piperaquine susceptibility assays do not correlate with treatment outcome. A novel assay using a pharmacologically relevant piperaquine dose/time exposure was designed and its relevance explored in retrospective and prospective studies.METHODS:The piperaquine survival assay (PSA) exposed parasites to 200 nM piperaquine for 48 hours and monitored survival 24 hours later. The retrospective study tested 32 culture-adapted, C580Y-K13 mutant parasites collected at enrolment from patients treated with a 3-day course of dihydroartemisinin-piperaquine and having presented or not with a recrudescence at day 42 (registered ACTRN12615000793516). The prospective study assessed ex vivo PSA survival rate alongside K13 polymorphism of isolates collected from patients enrolled in an open-label study with dihydroartemisinin-piperaquine for uncomplicated P. falciparum malaria in Cambodia (registered ACTRN12615000696594).RESULTS:All parasites from recrudescent cases had in vitro or ex vivo PSA survival rates ≥10%, a relevant cut-off value for piperaquine-resistance. Ex vivo PSA survival rates were higher for recrudescent than non-recrudescent cases (39.2% vs. 0.17%, P <1 × 10(-7)). Artemisinin-resistant K13 mutants with ex vivo PSA survival rates ≥10% were associated with 32-fold higher risk of recrudescence (95% CI, 4.5-224; P = 0.0005).CONCLUSION:PSA adequately captures the piperaquine resistance/recrudescence phenotype, a mainstay to identify molecular marker(s) and evaluate efficacy of alternative drugs. Combined ex vivo PSA and K13 genotyping provides a convenient monitor for both artemisinin and piperaquine resistance where dihydroartemisinin-piperaquine is used.
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