Baculovirus expression and diagnostic utility of the glycoprotein E of bovine herpesvirus-1.1 Egyptian strain "Abu-Hammad"

J Virol Methods. 2013 Jul;191(1):33-40. doi: 10.1016/j.jviromet.2013.03.024. Epub 2013 Apr 1.

Abstract

A recombinant baculovirus construct expressing glycoprotein E (gE) of the Egyptian BoHV-1.1 Abu-Hammad strain (rBac/gE-AbuH) was generated and characterized. The recombinant gE (rgE) secreting protein in culture medium of infected insect cells was used as a coating antigen in an indirect enzyme-linked immunosorbent assay (ELISA) to test its utility for detection of antibody against gE of BoHV-1. Indirect gE-ELISA was compared to standard virus neutralization test and commercial blocking gE-ELISA for detection of anti-gE antibody in a panel of bovine sera. Antibody titers estimated by both ELISAs were closely correlated with those determined by virus neutralization test. In conclusion, the developed indirect gE-ELISA was a reliable candidate for inexpensive detection of anti-gE antibody in control and experimental bovine sera with high specificity and sensitivity. Moreover, it emphasized the diagnostic utility of gE based ELISAs to distinguish cattle infected with BoHV-1 from those vaccinated with the gE negative mutants.

Publication types

  • Evaluation Study

MeSH terms

  • Animals
  • Antibodies, Viral / blood*
  • Baculoviridae / genetics
  • Cattle
  • Cattle Diseases / diagnosis*
  • Cell Line
  • Clinical Laboratory Techniques / methods*
  • Culture Media / chemistry
  • Enzyme-Linked Immunosorbent Assay / methods
  • Genetic Vectors
  • Herpesviridae Infections / diagnosis
  • Herpesviridae Infections / veterinary*
  • Herpesvirus 1, Bovine / immunology*
  • Insecta
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Sensitivity and Specificity
  • Veterinary Medicine / methods*
  • Viral Proteins* / genetics
  • Viral Proteins* / isolation & purification
  • Virology / methods

Substances

  • Antibodies, Viral
  • Culture Media
  • Recombinant Proteins
  • Viral Proteins
  • bovine herpesvirus type-1 glycoproteins