title: Analytical Validation of the ReEBOV Antigen Rapid Test for Point-of-Care Diagnosis of Ebola Virus Infection.
abstract: Ebola virus disease (EVD) is a severe viral illness caused by Ebola virus (EBOV). The 2013-2016 EVD outbreak in West Africa is the largest recorded, with >11 000 deaths. Development of the ReEBOV Antigen Rapid Test (ReEBOV RDT) was expedited to provide a point-of-care test for suspected EVD cases.
  
  
  
  Recombinant EBOV viral protein 40 antigen was used to derive polyclonal antibodies for RDT and enzyme-linked immunosorbent assay development. ReEBOV RDT limits of detection (LOD), specificity, and interference were analytically validated on the basis of Food and Drug Administration (FDA) guidance.
  
  
  
  The ReEBOV RDT specificity estimate was 95% for donor serum panels and 97% for donor whole-blood specimens. The RDT demonstrated sensitivity to 3 species of Ebolavirus (Zaire ebolavirus, Sudan ebolavirus, and Bundibugyo ebolavirus) associated with human disease, with no cross-reactivity by pathogens associated with non-EBOV febrile illness, including malaria parasites. Interference testing exhibited no reactivity by medications in common use. The LOD for antigen was 4.7 ng/test in serum and 9.4 ng/test in whole blood. Quantitative reverse transcription-polymerase chain reaction testing of nonhuman primate samples determined the range to be equivalent to 3.0 × 105-9.0 × 108genomes/mL.
  
  
  
  The analytical validation presented here contributed to the ReEBOV RDT being the first antigen-based assay to receive FDA and World Health Organization emergency use authorization for this EVD outbreak, in February 2015.
authors: Galveston National Laboratory, University of Texas Medical Branch.
authors: Robert W
authors: RW
authors: Cross
authors: Corgenix, Broomfield, Colorado Department of Microbiology and Immunology Zalgen Labs, Germantown, Maryland.
authors: Matthew L
authors: ML
authors: Boisen
authors: Corgenix, Broomfield, Colorado.
authors: Molly M
authors: MM
authors: Millett
authors: Corgenix, Broomfield, Colorado Zalgen Labs, Germantown, Maryland.
authors: Diana S
authors: DS
authors: Nelson
authors: Corgenix, Broomfield, Colorado.
authors: Darin
authors: D
authors: Oottamasathien
authors: Department of Microbiology and Immunology.
authors: Jessica N
authors: JN
authors: Hartnett
authors: Corgenix, Broomfield, Colorado.
authors: Abigal B
authors: AB
authors: Jones
authors: Lassa Fever Program, Kenema Government Hospital Ministry of Health and Sanitation.
authors: Augustine
authors: A
authors: Goba
authors: Lassa Fever Program, Kenema Government Hospital Ministry of Health and Sanitation Eastern Polytechnic Institute, Kenema, Sierra Leone.
authors: Mambu
authors: M
authors: Momoh
authors: Lassa Fever Program, Kenema Government Hospital Ministry of Health and Sanitation Eastern Polytechnic Institute, Kenema, Sierra Leone.
authors: Mohamed
authors: M
authors: Fullah
authors: The Scripps Research Institute, La Jolla, California.
authors: Zachary A
authors: ZA
authors: Bornholdt
authors: The Scripps Research Institute, La Jolla, California.
authors: Marnie L
authors: ML
authors: Fusco
authors: The Scripps Research Institute, La Jolla, California.
authors: Dafna M
authors: DM
authors: Abelson
authors: The Scripps Research Institute, La Jolla, California.
authors: Shunichiro
authors: S
authors: Oda
authors: Corgenix, Broomfield, Colorado.
authors: Bethany L
authors: BL
authors: Brown
authors: Corgenix, Broomfield, Colorado.
authors: Ha
authors: H
authors: Pham
authors: Zalgen Labs, Germantown, Maryland.
authors: Megan M
authors: MM
authors: Rowland
authors: Galveston National Laboratory, University of Texas Medical Branch.
authors: Krystle N
authors: KN
authors: Agans
authors: Galveston National Laboratory, University of Texas Medical Branch.
authors: Joan B
authors: JB
authors: Geisbert
authors: Zalgen Labs, Germantown, Maryland.
authors: Megan L
authors: ML
authors: Heinrich
authors: Autoimmune Technologies, New Orleans, Louisiana.
authors: Peter C
authors: PC
authors: Kulakosky
authors: School of Public Health and Tropical Medicine, Tulane University.
authors: Jeffrey G
authors: JG
authors: Shaffer
authors: Section of Infectious Diseases, Department of Pediatrics, School of Medicine.
authors: John S
authors: JS
authors: Schieffelin
authors: Ministry of Health and Sanitation.
authors: Brima
authors: B
authors: Kargbo
authors: Lassa Fever Program, Kenema Government Hospital Ministry of Health and Sanitation.
authors: Momoh
authors: M
authors: Gbetuwa
authors: Lassa Fever Program, Kenema Government Hospital University of Sierra Leone, Freetown.
authors: Sahr M
authors: SM
authors: Gevao
authors: Autoimmune Technologies, New Orleans, Louisiana.
authors: Russell B
authors: RB
authors: Wilson
authors: The Scripps Research Institute, La Jolla, California.
authors: Erica Ollmann
authors: EO
authors: Saphire
authors: Corgenix, Broomfield, Colorado.
authors: Kelly R
authors: KR
authors: Pitts
authors: Lassa Fever Program, Kenema Government Hospital Ministry of Health and Sanitation.
authors: Sheik Humarr
authors: SH
authors: Khan
authors: Lassa Fever Program, Kenema Government Hospital Ministry of Health and Sanitation.
authors: Donald S
authors: DS
authors: Grant
authors: Galveston National Laboratory, University of Texas Medical Branch.
authors: Thomas W
authors: TW
authors: Geisbert
authors: Zalgen Labs, Germantown, Maryland.
authors: Luis M
authors: LM
authors: Branco
authors: Department of Microbiology and Immunology Zalgen Labs, Germantown, Maryland.
authors: Robert F
authors: RF
authors: Garry
mesh_terms: D000354:Africa, Western / Q000453:epidemiology; D000818:Animals; D000956:Antigens, Viral / Q000097:blood*; D004196:Disease Outbreaks; D029043:Ebolavirus / Q000276:immunology* / Q000302:isolation & purification; D004797:Enzyme-Linked Immunosorbent Assay; D019142:Hemorrhagic Fever, Ebola / Q000175:diagnosis* / Q000821:virology; D006801:Humans; D007118:Immunoassay; D057230:Limit of Detection; D019095:Point-of-Care Systems; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D014763:Viral Matrix Proteins / Q000097:blood*
chemical_list: D000956:Antigens, Viral; D011933:Reagent Kits, Diagnostic; C080981:VP40 protein, virus; D014763:Viral Matrix Proteins
keywords: Ebola, point of care; diagnostic; lateral flow immunoassay