Figure 2. Detection of pestivirus E2 protein with monoclonal antibody 6A5 in investigation of piglets with congenital tremor (CT) on a farm in southeastern Austria, 2015. A) Western blot analysis of cells infected with novel virus provisionally termed Linda virus. Total protein of SK-6 cells infected with Linda virus and MDBK cells infected with BVDV-1 (strain NADL) was probed with the pestivirus E2-specific antibody 6A5. The apparent molecular mass of monomeric E2 (LV 50 kDa and BVDV-1 55 kDa) shows that Linda virus E2 has a lower molecular weight than BVDV-1 E2 as a result of fewer N-linked glycosylation sites. In contrast, the mass of E1-E2 heterodimers is comparable (≈70 kDa in Linda virus and BVDV-1), because Linda virus E1 has an additional N-linked glycosylation site. B, C) Focus size of Linda virus 48 hours after infection of SK-6 cells (B) and MDBK cells (C) (original magnification ×20). D, E) Detection of pestiviral E2 within neuronal tissue of Linda virus–positive, congenital tremor–affected piglets showing positive signals in neurons of the nucleus of the trigeminal nerve (D) (original magnification ×10) and within glial cells in the cerebellar white matter (E) (original magnification ×20). BVDV, bovine viral diarrhea virus.