Detection of Zaire ebolavirus in swine: Assay development and optimization

B. S. Pickering, B. Collignon, G. Smith, P. Marszal, G. Kobinger, H. M. Weingartl

Research output: Contribution to journalArticlepeer-review

Abstract

Ebolaviruses (family Filoviridae, order Mononegavirales) cause often fatal, haemorrhagic fever in primates including humans. Pigs have been identified as a species susceptible to Reston ebolavirus (RESTV) infection, with indicated transmission to humans in the Philippines; however, their role during Ebola outbreaks in Africa needs to be clarified. To perform surveillance studies, detection of ebolavirus requires a prerequisite validation of viral RNA and antibody detection methods in swine samples. These diagnostic tests also need to be suitable for deployment to low-level containment laboratories. In this study, we developed a set of tests for detection of antibodies against Zaire ebolavirus (EBOV) in swine. Recombinant EBOV nucleoprotein was produced using a baculovirus expression system for indirect ELISA development. Evaluation of this assay was performed using laboratory and field samples, achieving a diagnostic specificity of 99%. Importantly, the indirect ELISA was able to detect antibodies to EBOV at 7 dpi, 3 days earlier than virus neutralization tests (VNT). The format of the VNT in this work was modified to a microtitre plaque reduction neutralization assay (miPRNT) complemented with immunostaining to provide a more rapid and highly specific assay. Finally, a confirmatory immunoblot assay was generated to supplement the indirect ELISA results.

Original languageEnglish (US)
Pages (from-to)77-84
Number of pages8
JournalTransboundary and Emerging Diseases
Volume65
Issue number1
DOIs
StatePublished - Feb 2018
Externally publishedYes

Keywords

  • ELISA
  • Ebola virus
  • baculovirus
  • diagnostics
  • plaque immunostaining
  • serology

ASJC Scopus subject areas

  • General Immunology and Microbiology
  • General Veterinary

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