title: The Malaria TaqMan Array Card Includes 87 Assays for Plasmodium falciparum Drug Resistance, Identification of Species, and Genotyping in a Single Reaction.
abstract: Antimalarial drug resistance exacerbates the global disease burden and complicates eradication efforts. To facilitate the surveillance of resistance markers in countries of malaria endemicity, we developed a suite of TaqMan assays for known resistance markers and compartmentalized them into a single array card (TaqMan array card, TAC). We included 87 assays for species identification, for the detection ofPlasmodium falciparummutations associated with chloroquine, atovaquone, pyrimethamine, sulfadoxine, and artemisinin resistance, and for neutral single nucleotide polymorphism (SNP) genotyping. Assay performance was first optimized using DNA from common laboratory parasite lines and plasmid controls. The limit of detection was 0.1 to 10 pg of DNA and yielded 100% accuracy compared to sequencing. The tool was then evaluated on 87 clinical blood samples from around the world, and the malaria TAC once again achieved 100% accuracy compared to sequencing and in addition detected the presence of mixed infections in clinical samples. With its streamlined protocol and high accuracy, this malaria TAC should be a useful tool for large-scale antimalarial resistance surveillance.
authors.affiliation: Division of Infectious Diseases and International Health, Department of Medicine, University of Virginia, Charlottesville, Virginia, USA.
authors.forename: Suporn
authors.initials: S
authors.lastname: Pholwat
authors.affiliation: Division of Infectious Diseases and International Health, Department of Medicine, University of Virginia, Charlottesville, Virginia, USA.
authors.forename: Jie
authors.initials: J
authors.lastname: Liu
authors.affiliation: Division of Infectious Diseases and International Health, Department of Medicine, University of Virginia, Charlottesville, Virginia, USA.
authors.forename: Suzanne
authors.initials: S
authors.lastname: Stroup
authors.affiliation: Department of Medicine, Division of Allergy and Infectious Diseases, University of Washington, Seattle, Washington, USA.
authors.forename: Shevin T
authors.initials: ST
authors.lastname: Jacob
authors.affiliation: Department of Community Health, Masaka Regional Referral Hospital, Masaka, Uganda.
authors.forename: Patrick
authors.initials: P
authors.lastname: Banura
authors.affiliation: Division of Infectious Diseases and International Health, Department of Medicine, University of Virginia, Charlottesville, Virginia, USA.
authors.forename: Christopher C
authors.initials: CC
authors.lastname: Moore
authors.affiliation: Division of Malaria Research, Institute for Global Health, University of Maryland School of Medicine, Baltimore, Maryland, USA.|National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Shanghai, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Huang
authors.affiliation: Division of Malaria Research, Institute for Global Health, University of Maryland School of Medicine, Baltimore, Maryland, USA.
authors.forename: Miriam K
authors.initials: MK
authors.lastname: Laufer
authors.affiliation: Division of Infectious Diseases and International Health, Department of Medicine, University of Virginia, Charlottesville, Virginia, USA.
authors.forename: Eric
authors.initials: E
authors.lastname: Houpt
authors.affiliation: Division of Infectious Diseases and International Health, Department of Medicine, University of Virginia, Charlottesville, Virginia, USA jlg5fw@virginia.edu.|Department of Biology, University of Virginia, Charlottesville, Virginia, USA.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Guler
mesh_terms: D000962:Antimalarials / Q000494:pharmacology*; D037621:Artemisinins / Q000494:pharmacology; D053626:Atovaquone / Q000494:pharmacology; D002738:Chloroquine / Q000494:pharmacology; D004351:Drug Resistance; D062665:Epidemiological Monitoring; D060005:Genotyping Techniques; D006801:Humans; D016778:Malaria, Falciparum / Q000188:drug therapy* / Q000469:parasitology; D010963:Plasmodium falciparum / Q000145:classification / Q000187:drug effects / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D011739:Pyrimethamine / Q000494:pharmacology; D011933:Reagent Kits, Diagnostic; D013413:Sulfadoxine / Q000494:pharmacology
chemical_list: D000962:Antimalarials; D037621:Artemisinins; D011933:Reagent Kits, Diagnostic; D013413:Sulfadoxine; D002738:Chloroquine; C031327:artemisinin; D053626:Atovaquone; D011739:Pyrimethamine
keywords: TaqMan PCR; antimalarial resistance; malaria; mutation; surveillance tool