title: High-throughput analysis of antimalarial susceptibility data by the WorldWide Antimalarial Resistance Network (WWARN) in vitro analysis and reporting tool.
abstract: Assessment of in vitro susceptibility is a fundamental component of antimalarial surveillance studies, but wide variations in the measurement of parasite growth and the calculation of inhibitory constants make comparisons of data from different laboratories difficult. Here we describe a Web-based, high-throughput in vitro analysis and reporting tool (IVART) generating inhibitory constants for large data sets. Fourteen primary data sets examining laboratory-determined susceptibility to artemisinin derivatives and artemisinin combination therapy partner drugs were collated from 11 laboratories. Drug concentrations associated with half-maximal inhibition of growth (IC50s) were determined by a modified sigmoid Emax model-fitting algorithm, allowing standardized analysis of 7,350 concentration-inhibition assays involving 1,592 isolates. Examination of concentration-inhibition data revealed evidence of apparent paradoxical growth at high concentrations of nonartemisinin drugs, supporting amendment of the method for calculating the maximal drug effect in each assay. Criteria for defining more-reliable IC50s based on estimated confidence intervals and growth ratios improved correlation coefficients for the drug pairs mefloquine-quinine and chloroquine-desethylamodiaquine in 9 of 11 and 8 of 8 data sets, respectively. Further analysis showed that maximal drug inhibition was higher for artemisinins than for other drugs, particularly in ELISA (enzyme-linked immunosorbent assay)-based assays, a finding consistent with the earlier onset of action of these drugs in the parasite life cycle. This is the first high-throughput analytical approach to apply consistent constraints and reliability criteria to large, diverse antimalarial susceptibility data sets. The data also illustrate the distinct biological properties of artemisinins and underline the need to apply more sensitive approaches to assessing in vitro susceptibility to these drugs.
authors.affiliation: WorldWide Antimalarial Resistance Network (WWARN), Centre for Tropical Medicine, Nuffield Department of Clinical Medicine, University of Oxford, Churchill Hospital, Oxford, United Kingdom. charlie@tropmedres.ac
authors.forename: Charles J
authors.initials: CJ
authors.lastname: Woodrow
authors.forename: Sabina
authors.initials: S
authors.lastname: Dahlström
authors.forename: Richard
authors.initials: R
authors.lastname: Cooksey
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Flegg
authors.forename: Hervé
authors.initials: H
authors.lastname: Le Nagard
authors.forename: France
authors.initials: F
authors.lastname: Mentré
authors.forename: Claribel
authors.initials: C
authors.lastname: Murillo
authors.forename: Didier
authors.initials: D
authors.lastname: Ménard
authors.forename: François
authors.initials: F
authors.lastname: Nosten
authors.forename: Kanlaya
authors.initials: K
authors.lastname: Sriprawat
authors.forename: Lise
authors.initials: L
authors.lastname: Musset
authors.forename: Neils B
authors.initials: NB
authors.lastname: Quashie
authors.forename: Pharath
authors.initials: P
authors.lastname: Lim
authors.forename: Rick M
authors.initials: RM
authors.lastname: Fairhurst
authors.forename: Sam L
authors.initials: SL
authors.lastname: Nsobya
authors.forename: Veronique
authors.initials: V
authors.lastname: Sinou
authors.forename: Harald
authors.initials: H
authors.lastname: Noedl
authors.forename: Bruno
authors.initials: B
authors.lastname: Pradines
authors.forename: Jacob D
authors.initials: JD
authors.lastname: Johnson
authors.forename: Philippe J
authors.initials: PJ
authors.lastname: Guerin
authors.forename: Carol H
authors.initials: CH
authors.lastname: Sibley
authors.forename: Jacques
authors.initials: J
authors.lastname: Le Bras
mesh_terms: D000655:Amodiaquine / Q000031:analogs & derivatives / Q000494:pharmacology; D000962:Antimalarials / Q000494:pharmacology*; D037621:Artemisinins / Q000494:pharmacology*; D057205:Automation, Laboratory; D002738:Chloroquine / Q000494:pharmacology; D004351:Drug Resistance; D004359:Drug Therapy, Combination; D057166:High-Throughput Screening Assays; D020407:Internet; D016778:Malaria, Falciparum / Q000188:drug therapy* / Q000469:parasitology; D015767:Mefloquine / Q000494:pharmacology; D021261:Parasitic Sensitivity Tests; D010963:Plasmodium falciparum / Q000187:drug effects*; D011803:Quinine / Q000494:pharmacology
chemical_list: D000962:Antimalarials; D037621:Artemisinins; D000655:Amodiaquine; C047386:desethylamodiaquine; D002738:Chloroquine; C031327:artemisinin; D011803:Quinine; D015767:Mefloquine