Glycoprotein gE blocking ELISAs to differentiate between Aujeszky's disease-vaccinated and infected animals

• Published in: Journal of virological methods 1997-04, Vol.65 (1), p.83-94
• Main Authors: Morenkov, O.S., Sobko, Yu.A., Panchenko, O.A.
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Monoclonal - immunology; Antibodies, Monoclonal - isolation & purification; ANTICORPS MONOCLONAL; ANTICUERPOS MONOCLONALES; AUJESZKY VIRUS; Aujeszky's disease virus; CERDO; ELISA; Enzyme-Linked Immunosorbent Assay - methods; Enzyme-Linked Immunosorbent Assay - veterinary; gE-ELISA; GLICOPROTEINAS; Glycoprotein E; GLYCOPROTEINE; GLYCOPROTEINS; Glycoproteins - analysis; Glycoproteins - chemistry; Glycoproteins - immunology; Herpesviridae Infections - immunology; Herpesviridae Infections - veterinary; INFECCION; INFECTION; MONOCLONAL ANTIBODIES; PORCIN; Pseudorabies - virology; Sensitivity and Specificity; Serological differentiation; SWINE; TEST ELISA; VACCINATION; Vaccination - veterinary; VACUNACION; VIRUS D'AUJESZKY; VIRUS DE AUJESZKY
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/S0166-0934(96)02171-4

Three blocking ELISAs for the detection of antibodies against glycoprotein E (gE) of Aujeszky's disease virus (ADV) in sera (an indirect blocking ELISA (gE-iELISA), a direct blocking ELISA (gE-dELISA) and a two-site ‘sandwich’ blocking ELISA (gE-sELISA)) have been developed. The gE-ELISAs are based on monoclonal antibodies (mAbs) directed to gE and detect gE-specific antibodies in sera by blocking the binding of mAbs to one (in the gE-iELISA and the gE-dELISA) or two (in the gE-sELISA) epitopes of gE. From a panel of thirteen gE-specific mAbs, mAbs, their conjugates and the combination coating mAb/conjugate that optimally detect anti-gE antibodies in the assays were selected. Sera from uninfected unvaccinated swine or swine vaccinated against different swine viral disorders were negative by three gE-ELISAs, the blocking gB-ELISA, and VNT. Swine vaccinated with gE-negative vaccine were seropositive in the gB-ELISA and VNT but were seronegative by three gE-ELISAs. Infected unvaccinated swine, infected swine vaccinated with gE-negative vaccine, and swine vaccinated with gE-positive vaccine were detected as seropositive by three gE-ELISAs as well as in the gB-ELISA. The gE-dELISA and the gE-sELISA proved to be specific and the most sensitive and reliable assays to distinguish ADV-infected swine from those vaccinated with gE-negative vaccine. These two gE-ELISAs were at least as sensitive as the gB-ELISA for detecting ADV-specific antibodies.