title: Development and validation of a genotype 3 recombinant protein-based immunoassay for hepatitis E virus serology in swine.
abstract: Hepatitis E virus (HEV) is classified within the family Hepeviridae, genus Hepevirus. HEV genotype 3 (Gt3) infections are endemic in pigs in Western Europe and in North and South America and cause zoonotic infections in humans. Several serological assays to detect HEV antibodies in pigs have been developed, at first mainly based on HEV genotype 1 (Gt1) antigens. To develop a sensitive HEV Gt3 ELISA, a recombinant baculovirus expression product of HEV Gt3 open reading frame-2 was produced and coated onto polystyrene ELISA plates. After incubation of porcine sera, bound HEV antibodies were detected with anti-porcine anti-IgG and anti-IgM conjugates. For primary estimation of sensitivity and specificity of the assay, sets of sera were used from pigs experimentally infected with HEV Gt3. For further validation of the assay and to set the cutoff value, a batch of 1100 pig sera was used. All pig sera were tested using the developed HEV Gt3 assay and two other serologic assays based on HEV Gt1 antigens. Since there is no gold standard available for HEV antibody testing, further validation and a definite setting of the cutoff of the developed HEV Gt3 assay were performed using a statistical approach based on Bayes' theorem. The developed and validated HEV antibody assay showed effective detection of HEV-specific antibodies. This assay can contribute to an improved detection of HEV antibodies and enable more reliable estimates of the prevalence of HEV Gt3 in swine in different regions.
authors.affiliation: Central Veterinary Institute, Wageningen University and Research Centre, Lelystad, The Netherlands.
authors.forename: W H M
authors.initials: WH
authors.lastname: van der Poel
authors.affiliation: Animal Health Laboratory UMR 1161, Agence Nationale de Sécurité Sanitaire de l'Alimentation, de l'Environnement et du Travail, Maisons-Alfort, France.
authors.forename: N
authors.initials: N
authors.lastname: Pavio
authors.affiliation: Central Veterinary Institute, Wageningen University and Research Centre, Lelystad, The Netherlands.
authors.forename: J
authors.initials: J
authors.lastname: van der Goot
authors.affiliation: Central Veterinary Institute, Wageningen University and Research Centre, Lelystad, The Netherlands.
authors.forename: M
authors.initials: M
authors.lastname: van Es
authors.affiliation: Departament de Sanitat i d'Anatomia Animals, Centre de Recerca en Sanitat Animal, Universitat Autònoma de Barcelona, Institut de Recerca i Tecnologia Agroalimentaries, Bellaterra, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Martin
authors.affiliation: Biometris, Wageningen University and Research Centre, Wageningen, The Netherlands.
authors.forename: B
authors.initials: B
authors.lastname: Engel
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D016367:Baculoviridae; D001499:Bayes Theorem; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors; D005838:Genotype; D016751:Hepatitis E / Q000097:blood / Q000662:veterinary*; D016752:Hepatitis E virus / Q000145:classification / Q000302:isolation & purification*; D006524:Hepatitis, Viral, Animal / Q000175:diagnosis*; D016366:Open Reading Frames; D011994:Recombinant Proteins; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012698:Serologic Tests; D013552:Swine / Q000821:virology*
chemical_list: D000914:Antibodies, Viral; D011994:Recombinant Proteins