RESUMO
Three modifications of quantitative enzyme immunoassay (EIA) for assessment of Aujeszky's disease virus (ADV) in culture medium are compared, in which monoclonal antibodies (MAB) to ADV GH glycoprotein were used: nonconcurrent two-site "sandwich" EIA on the basis of two different MABs, indirect concurrent EIA, and direct concurrent EIA. MABs fit best of all for use in every of EIA modifications were selected. Conditions of the assays were optimized. Concurrent EIAs were 10-20 times less sensitive than sandwich EIA. The developed sandwich EIA permitted ADV assay in preparations of both live and formaldehyde-inactivated virus.
Assuntos
Anticorpos Monoclonais , Pseudorraiva/diagnóstico , Animais , Chlorocebus aethiops , Meios de Cultura , Herpesvirus Suídeo 1/imunologia , Técnicas Imunoenzimáticas , Células VeroRESUMO
A panel of 24 different monoclonal antibodies (MAB) to Aujeszky's disease virus (ADV) proteins was prepared. Seven MABs were directed to ADV glycoprotein GII (6 MABs to GIIc chain and 1 MAB to GIIb chain). GII epitope mapping with the resultant MABs was carried out. Four ADV GII epitopes were detected: epitopes A, B, and C located on GIIc chain and epitope D located on GIIb chain. Epitope B overlaps epitopes A and C which do not overlap each other, epitope D does not overlap other epitopes. Epitopes A, B, and C are located in a restricted area of GIIc chain which seems to be one of GII immunodominant regions. The detected epitopes are mainly conformational, for they are sensitive to denaturation. Of all MABs obtained more than 30% were directed to ADV GII. This indicates that GII is one of the most important ADV antigens. All the seven MABs to GII interacted with all tested ADV strains (K. Arsky, VGNKI, BUK, and NIA-4), this being in line with the data on a highly conservative nature of ADV GII.