title: Development of a sandwich ELISA for the detection of bovine herpesvirus type 1.
abstract: OBJECTIVE
  To develop a standard enzyme-linked immunosorbent assay (ELISA) for the detection of bovine herpesvirus type 1 (BHV-1).
  
  
  METHODS
  The assay was based on hyperimmune rabbit and guinea pig antisera raised against purified BHV-1. Polyethylene glycol precipitation and sucrose density gradient methods were adopted for viral concentration and purification. Antisera were raised using Freund's adjuvant followed by extraction of IgG of high purity.
  
  
  RESULTS
  Optimum antisera dilutions as determined by titrations were chosen as 1:4 000, whereas the conjugate was used at 1:2 000 dilution. Using 95 clinical specimens, the ELISA test showed a sensitivity and specificity of 91.90 % and 93.10 %, respectively when compared to PCR. The cut-off value was fixed at 0.15 (A(490)) and a P/N ratio of >1.30 indicated a significant positive reaction.
  
  
  CONCLUSIONS
  The results have demonstrated that this ELISA could efficiently detect BHV-1 and can be used as an important diagnostic tool.
authors.affiliation: Department of Epidemiology and Vety. Preventive medicine, College of Veterinary Sciences, Veterinary University, Mathura (UP), India.
authors.forename: Shanaz
authors.initials: S
authors.lastname: Bashir
authors.forename: Rashmi
authors.initials: R
authors.lastname: Singh
authors.forename: Barkha
authors.initials: B
authors.lastname: Sharma
authors.forename: Sharad K
authors.initials: SK
authors.lastname: Yadav
mesh_terms: D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006168:Guinea Pigs; D007242:Herpesvirus 1, Bovine / Q000302:isolation & purification*; D011817:Rabbits; D012680:Sensitivity and Specificity