title: Development, Evaluation, and Integration of a Quantitative Reverse-Transcription Polymerase Chain Reaction Diagnostic Test for Ebola Virus on a Molecular Diagnostics Platform.
abstract: The 2013-2016 Ebola epidemic in West Africa resulted in accelerated development of rapid diagnostic tests for emergency outbreak preparedness. We describe the development and evaluation of the Idylla™ prototype Ebola virus test, a fully automated sample-to-result molecular diagnostic test for rapid detection of Zaire ebolavirus (EBOV) and Sudan ebolavirus (SUDV).
  
  
  
  The Idylla™ prototype Ebola virus test can simultaneously detect EBOV and SUDV in 200 µL of whole blood. The sample is directly added to a disposable cartridge containing all reagents for sample preparation, RNA extraction, and amplification by reverse-transcription polymerase chain reaction analysis. The performance was evaluated with a variety of sample types, including synthetic constructs and whole blood samples from healthy volunteers spiked with viral RNA, inactivated virus, and infectious virus.
  
  
  
  The 95% limits of detection for EBOV and SUDV were 465 plaque-forming units (PFU)/mL (1010 copies/mL) and 324 PFU/mL (8204 copies/mL), respectively. In silico and in vitro analyses demonstrated 100% correct reactivity for EBOV and SUDV and no cross-reactivity with relevant pathogens. The diagnostic sensitivity was 97.4% (for EBOV) and 91.7% (for SUDV), the specificity was 100%, and the diagnostic accuracy was 95.9%.
  
  
  
  The Idylla™ prototype Ebola virus test is a fast, safe, easy-to-use, and near-patient test that meets the performance criteria to detect EBOV in patients with suspected Ebola.
authors.affiliation: Department of Clinical Sciences.
authors.forename: Lieselotte
authors.initials: L
authors.lastname: Cnops
authors.affiliation: Janssen Diagnostics, Janssen Pharmaceutica, Beerse Biocartis, Mechelen, Belgium.
authors.forename: Peter
authors.initials: P
authors.lastname: Van den Eede
authors.affiliation: National Institute of Allergy and Infectious Diseases Integrated Research Facility, National Institutes of Health, Frederick, Maryland.
authors.forename: James
authors.initials: J
authors.lastname: Pettitt
authors.affiliation: Department of Biomedical Sciences, Institute of Tropical Medicine, Antwerp.
authors.forename: Leo
authors.initials: L
authors.lastname: Heyndrickx
authors.affiliation: Department of Clinical Sciences.
authors.forename: Birgit
authors.initials: B
authors.lastname: De Smet
authors.affiliation: Department of Biomedical Sciences, Institute of Tropical Medicine, Antwerp.
authors.forename: Sandra
authors.initials: S
authors.lastname: Coppens
authors.affiliation: Janssen Diagnostics, Janssen Pharmaceutica, Beerse.
authors.forename: Ilse
authors.initials: I
authors.lastname: Andries
authors.affiliation: Janssen Diagnostics, Janssen Pharmaceutica, Beerse.
authors.forename: Theresa
authors.initials: T
authors.lastname: Pattery
authors.affiliation: Biocartis, Mechelen, Belgium.
authors.forename: Luc
authors.initials: L
authors.lastname: Van Hove
authors.affiliation: Biocartis, Mechelen, Belgium.
authors.forename: Geert
authors.initials: G
authors.lastname: Meersseman
authors.affiliation: Biocartis, Mechelen, Belgium.
authors.forename: Sari
authors.initials: S
authors.lastname: Van Den Herrewegen
authors.affiliation: Biocartis, Mechelen, Belgium.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Vergauwe
authors.affiliation: Biocartis, Mechelen, Belgium.
authors.forename: Rein
authors.initials: R
authors.lastname: Thijs
authors.affiliation: National Institute of Allergy and Infectious Diseases Integrated Research Facility, National Institutes of Health, Frederick, Maryland.
authors.forename: Peter B
authors.initials: PB
authors.lastname: Jahrling
authors.affiliation: Biocartis, Mechelen, Belgium.
authors.forename: David
authors.initials: D
authors.lastname: Nauwelaers
authors.affiliation: Department of Biomedical Sciences, Institute of Tropical Medicine, Antwerp.
authors.forename: Kevin K
authors.initials: KK
authors.lastname: Ariën
mesh_terms: D000354:Africa, Western / Q000453:epidemiology; D004196:Disease Outbreaks; D029043:Ebolavirus / Q000235:genetics / Q000302:isolation & purification*; D019142:Hemorrhagic Fever, Ebola / Q000175:diagnosis* / Q000453:epidemiology / Q000821:virology; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000295:instrumentation / Q000379:methods; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D012367:RNA, Viral
keywords: EBOV; Ebolavirus; RT-PCR; SUDV; diagnosis; outbreak