Development of Prototype Filovirus Recombinant Antigen Immunoassays

J Infect Dis. 2015 Oct 1;212 Suppl 2(Suppl 2):S359-67. doi: 10.1093/infdis/jiv353. Epub 2015 Jul 30.


Background: Throughout the 2014-2015 Ebola outbreak in West Africa, major gaps were exposed in the availability of validated rapid diagnostic platforms, protective vaccines, and effective therapeutic agents. These gaps potentiated the development of prototype rapid lateral flow immunodiagnostic (LFI) assays that are true point-of-contact platforms, for the detection of active Ebola infections in small blood samples.

Methods: Recombinant Ebola and Marburg virus matrix VP40 and glycoprotein (GP) antigens were used to derive a panel of monoclonal and polyclonal antibodies. Antibodies were tested using a multivariate approach to identify antibody-antigen combinations suitable for enzyme-linked immunosorbent assay (ELISA) and LFI assay development.

Results: Polyclonal antibodies generated in goats were superior reagents for capture and detection of recombinant VP40 in test sample matrices. These antibodies were optimized for use in antigen-capture ELISA and LFI assay platforms. Prototype immunoglobulin M (IgM)/immunoglobulin G (IgG) ELISAs were similarly developed that specifically detect Ebola virus-specific antibodies in the serum of experimentally infected nonhuman primates and in blood samples obtained from patients with Ebola from Sierra Leone.

Conclusions: The prototype recombinant Ebola LFI assays developed in these studies have sensitivities that are useful for clinical diagnosis of acute ebolavirus infections. The antigen-capture and IgM/IgG ELISAs provide additional confirmatory assay platforms for detecting VP40 and other ebolavirus-specific immunoglobulins.

Keywords: ELISA; Ebola; Ebola virus; filovirus; lateral flow immunodiagnostic; point-of-care testing.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Africa, Western
  • Animals
  • Antibodies, Viral / blood
  • Antibodies, Viral / immunology
  • Antigens, Viral / immunology*
  • Cross Reactions / immunology
  • Ebolavirus / immunology
  • Enzyme-Linked Immunosorbent Assay / methods
  • Filoviridae / immunology*
  • Hemorrhagic Fever, Ebola / blood
  • Hemorrhagic Fever, Ebola / immunology
  • Hemorrhagic Fever, Ebola / virology
  • Humans
  • Immunoassay / methods*
  • Immunoglobulin G / blood
  • Immunoglobulin G / immunology
  • Immunoglobulin M / blood
  • Immunoglobulin M / immunology
  • Marburg Virus Disease / blood
  • Marburg Virus Disease / immunology
  • Marburg Virus Disease / virology
  • Marburgvirus / immunology
  • Sierra Leone


  • Antibodies, Viral
  • Antigens, Viral
  • Immunoglobulin G
  • Immunoglobulin M