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...

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Bibliographic Details
Published in:Journal of fish diseases 2004-03, Vol.27 (3), p.143-149
Main Authors: Jewhurst, V A, Todd, D, Rowley, H M, Walker, I W, Weston, J H, McLoughlin, M F, Graham, D A
Format: Article
Language:English
Subjects:
alphavirus
Alphavirus - immunology
Alphavirus - isolation & purification
Alphavirus Infections - diagnosis
Alphavirus Infections - veterinary
Animals
Antibodies, Monoclonal - immunology
Aquaculture - methods
Cells, Cultured
Fish Diseases - blood
Fish Diseases - diagnosis
Fish Diseases - virology
Fluorescent Antibody Technique
Immunoenzyme Techniques - methods
Immunoenzyme Techniques - veterinary
Oncorhynchus mykiss
Salmo salar
salmon pancreas disease
salmon pancreas disease virus
sleeping disease
sleeping disease virus
viraemia
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Summary: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 °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.
ISSN:0140-7775
1365-2761
DOI:10.1111/j.1365-2761.2004.00530.x