Detection and antigenic characterization of salmonid alphavirus isolates from sera obtained from farmed Atlantic salmon, Salmo salar L., and farmed rainbow trout, Oncorhynchus mykiss (Walbaum).
A simple method of detecting the presence of the salmonid alphaviruses (SAVs), salmon pancreas disease virus (SPDV) and sleeping disease virus (SDV), from serum samples is described. Using a 96-well tissue-culture plate format, test sera are diluted in medium and added to chinook salmon embryo (CHSE-214) cells. After incubation for 3 days at 15 degrees C, plates are fixed and stained using a monoclonal antibody (mAb)-based immunoperoxidase (IPX) detection system, and virus-infected cells are observed microscopically by white light. Application of this screening test, which is now used routinely in our laboratory in conjunction with an IPX-based virus neutralization (IPX-VN) test for detecting antibodies to SAVs, has resulted in the recovery of 12 additional isolates from salmon sera and four additional isolates from trout sera. A low level of antigenic variation was detected when these SAV isolates were investigated by indirect immunofluorescence using a panel of mAbs raised to reference SPDV and SDV isolates.
Department of Veterinary Science, The Queen's University of Belfast, Veterinary Sciences Division, Stormont, Belfast, UK.
V A
VA
Jewhurst
D
D
Todd
H M
HM
Rowley
I W
IW
Walker
J H
JH
Weston
M F
MF
McLoughlin
D A
DA
Graham
D000524:Alphavirus / Q000276:immunology / Q000302:isolation & purification*; D018354:Alphavirus Infections / Q000175:diagnosis / Q000662:veterinary*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D017756:Aquaculture / Q000379:methods*; D002478:Cells, Cultured; D005393:Fish Diseases / Q000097:blood / Q000175:diagnosis* / Q000821:virology*; D005455:Fluorescent Antibody Technique; D007124:Immunoenzyme Techniques / Q000379:methods / Q000662:veterinary*; D017686:Oncorhynchus mykiss; D020317:Salmo salar
D000911:Antibodies, Monoclonal