Analytical Validation of the ReEBOV Antigen Rapid Test for Point-of-Care Diagnosis of Ebola Virus Infection

J Infect Dis. 2016 Oct 15;214(suppl 3):S210-S217. doi: 10.1093/infdis/jiw293. Epub 2016 Aug 31.


Background: 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.

Methods: 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.

Results: 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 × 108 genomes/mL.

Conclusions: 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.

Keywords: Ebola, point of care; diagnostic; lateral flow immunoassay.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Africa, Western / epidemiology
  • Animals
  • Antigens, Viral / blood*
  • Disease Outbreaks*
  • Ebolavirus / immunology*
  • Ebolavirus / isolation & purification
  • Enzyme-Linked Immunosorbent Assay
  • Hemorrhagic Fever, Ebola / diagnosis*
  • Hemorrhagic Fever, Ebola / virology
  • Humans
  • Immunoassay
  • Limit of Detection
  • Point-of-Care Systems*
  • Reagent Kits, Diagnostic
  • Sensitivity and Specificity
  • Viral Matrix Proteins / blood*


  • Antigens, Viral
  • Reagent Kits, Diagnostic
  • VP40 protein, virus
  • Viral Matrix Proteins