title: Detection of rainbow trout antibodies against viral haemorrhagic septicaemia virus (VHSV) by neutralisation test is highly dependent on the virus isolate used.
abstract: Three serological tests, enzyme linked immunosorbent assay (ELISA), 50% plaque neutralisation test (50%PNT) and Western blotting (WB), were used to detect antibodies against viral haemorrhagic septicaemia virus (VHSV) in 50 rainbow trout broodstock from a rainbow trout farm endemically infected with VHS but with no clinical signs of infection. When the sera were examined by 50%PNT using the VHSV reference isolate DK-F1 or the heat attenuated DK-F25 mutant strain, no neutralizing antibodies were found. In contrast, when one of the virus isolates from the farm (homologous virus) was used in the 50%PNT, 90% of the fish were found to be positive. By examining a panel of different VHSV isolates in 50%PNT, it was demonstrated that the virus isolate used as test antigen could significantly affect the sensitivity and titre determination in 50%PNT for detection of rainbow trout antibodies against VHSV. When the sera were examined for the presence of VHSV antibodies by ELISA or WB, 61% were found to be positive. When conducting WB analysis, the viral glycoprotein was the protein most frequently recognized, followed by the viral nucleoprotein.
authors: National Veterinary Institute, Technical University of Denmark, Hangøvej 2, 8200 Arhus N, Denmark.
authors: J M
authors: JM
authors: Fregeneda-Grandes
authors: N J
authors: NJ
authors: Olesen
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D015153:Blotting, Western / Q000662:veterinary; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D031941:Hemorrhagic Septicemia, Viral / Q000276:immunology* / Q000821:virology; D009500:Neutralization Tests / Q000662:veterinary; D029141:Novirhabdovirus / Q000276:immunology* / Q000302:isolation & purification; D017686:Oncorhynchus mykiss; D012680:Sensitivity and Specificity
chemical_list: D000914:Antibodies, Viral