title: Detection of Zaire ebolavirus in swine: Assay development and optimization.
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.
authors.affiliation: National Centre for Foreign Animal Disease, Canadian Food Inspection Agency, Winnipeg, MB, Canada.
authors.forename: B S
authors.initials: BS
authors.lastname: Pickering
authors.affiliation: National Centre for Foreign Animal Disease, Canadian Food Inspection Agency, Winnipeg, MB, Canada.
authors.forename: B
authors.initials: B
authors.lastname: Collignon
authors.affiliation: National Centre for Foreign Animal Disease, Canadian Food Inspection Agency, Winnipeg, MB, Canada.
authors.forename: G
authors.initials: G
authors.lastname: Smith
authors.affiliation: National Centre for Foreign Animal Disease, Canadian Food Inspection Agency, Winnipeg, MB, Canada.
authors.forename: P
authors.initials: P
authors.lastname: Marszal
authors.affiliation: Department of Microbiology, Immunology, faculty of Medicine, Université Laval, Québec, QC, Canada.
authors.forename: G
authors.initials: G
authors.lastname: Kobinger
authors.affiliation: National Centre for Foreign Animal Disease, Canadian Food Inspection Agency, Winnipeg, MB, Canada.|Department of Medical Microbiology, Faculty of Medicine, University of Manitoba, Winnipeg, MB, Canada.
authors.forename: H M
authors.initials: HM
authors.lastname: Weingartl
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology*; D015023:Democratic Republic of the Congo / Q000453:epidemiology; D004196:Disease Outbreaks / Q000662:veterinary*; D029043:Ebolavirus / Q000276:immunology / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D019142:Hemorrhagic Fever, Ebola / Q000453:epidemiology / Q000662:veterinary* / Q000821:virology; D006801:Humans; D009500:Neutralization Tests / Q000662:veterinary; D009698:Nucleoproteins / Q000235:genetics; D010679:Philippines / Q000453:epidemiology; D012367:RNA, Viral / Q000032:analysis; D013552:Swine; D013553:Swine Diseases / Q000453:epidemiology* / Q000821:virology
chemical_list: D000914:Antibodies, Viral; D009698:Nucleoproteins; D012367:RNA, Viral
keywords: 
  ELISA
  ; Ebola virus; baculovirus; diagnostics; plaque immunostaining; serology