title: Development and validation of an indirect ELISA as a confirmatory test for surveillance of infectious bovine rhinotracheitis in vaccinated herds.
abstract: BACKGROUND
  Bovine herpesvirus 1 (BoHV1) is a member of the viral subfamily of Alphaherpesvirinae that infects various species, including cattle, sheep, and goats. The virus causes infectious bovine rhinotracheitis (IBR), which is included in a European list of diseases that may require control and eradication programs. The lack of confirmatory tests affects the validity of diagnostic tools, especially those used for vaccinated herds. In this study, we report the development and validation of an indirect enzyme-linked immunosorbent assay (ELISA) based on BoHV1 glycoprotein E, which was expressed as a secreted recombinant antigen in a mammalian cell system. The performance of the new rec-gE ELISA was compared with that of commercially available indirect and/or blocking ELISAs.
  
  
  RESULTS
  The sample set included blood sera from animals from IBR-positive farms, IBR-free farms, and marker-vaccinated farms. The indirect ELISA proposed in this study is based on antibody reactivity against BoHV1 gE, and showed high sensitivity and specificity (98.41 and 99.76 %, respectively).
  
  
  CONCLUSIONS
  The ELISA performed well, in terms of both its diagnostic sensitivity and specificity, and as a confirmatory methodology, and therefore should improve the diagnostic protocols used for IBR surveillance.
authors.affiliation: Department of Veterinary Science, University of Torino, Torino, Italy. luigi.bertolotti@unito.it.|, Present address: Largo Paolo Braccini, 2, 10095, Grugliasco, TO, Italy. luigi.bertolotti@unito.it.
authors.forename: Luigi
authors.initials: L
authors.lastname: Bertolotti
authors.affiliation: Department of Veterinary Science, University of Torino, Torino, Italy. elvira.muratore@unito.it.
authors.forename: Elvira
authors.initials: E
authors.lastname: Muratore
authors.affiliation: Department of Veterinary Science, University of Torino, Torino, Italy. chiara.nogarol@unito.it.
authors.forename: Chiara
authors.initials: C
authors.lastname: Nogarol
authors.affiliation: Istituto Zooprofilattico Sperimentale di Piemonte, Liguria e Valle d'Aosta, Torino, Italy. claudio.caruso@izsto.it.
authors.forename: Claudio
authors.initials: C
authors.lastname: Caruso
authors.affiliation: Istituto Zooprofilattico Sperimentale delle Venezie, Legnaro, Padova, Italy. LLucchese@izsvenezie.it.
authors.forename: Laura
authors.initials: L
authors.lastname: Lucchese
authors.affiliation: Department of Veterinary Science, University of Torino, Torino, Italy. margherita.profiti@unito.it.
authors.forename: Margherita
authors.initials: M
authors.lastname: Profiti
authors.affiliation: Department of Chemistry, University of Torino, Torino, Italy. laura.anfossi@unito.it.
authors.forename: Laura
authors.initials: L
authors.lastname: Anfossi
authors.affiliation: Istituto Zooprofilattico Sperimentale di Piemonte, Liguria e Valle d'Aosta, Torino, Italy. loretta.masoero@izsto.it.
authors.forename: Loretta
authors.initials: L
authors.lastname: Masoero
authors.affiliation: Istituto Zooprofilattico Sperimentale delle Venezie, Legnaro, Padova, Italy. snardelli@izsvenezie.it.
authors.forename: Stefano
authors.initials: S
authors.lastname: Nardelli
authors.affiliation: Department of Veterinary Science, University of Torino, Torino, Italy. sergio.rosati@unito.it.
authors.forename: Sergio
authors.initials: S
authors.lastname: Rosati
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000276:immunology; D002417:Cattle; D002418:Cattle Diseases / Q000517:prevention & control*; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D007242:Herpesvirus 1, Bovine / Q000276:immunology / Q000378:metabolism; D007241:Infectious Bovine Rhinotracheitis / Q000517:prevention & control*; D011159:Population Surveillance; D011994:Recombinant Proteins; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014764:Viral Proteins / Q000276:immunology; D014765:Viral Vaccines / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D011994:Recombinant Proteins; D014764:Viral Proteins; D014765:Viral Vaccines; C046998:bovine herpesvirus type-1 glycoproteins