A rapid immunoperoxidase-based virus neutralization assay for salmonid alphavirus used for a serological survey in Northern Ireland.
A modified virus neutralization (VN) assay was developed to replace an existing assay read on the presence or absence of virus-induced cytopathic effect (CPE). The modified assay used a monoclonal antibody to salmon pancreas disease virus as the first layer of an immunoperoxidase (IPX)-based immunostaining technique to detect viral growth. The IPX-based VN assay required only 3 days to perform, and the adoption of a 96-well microtitre format facilitated a high throughput of samples requiring small volumes of serum, cells and virus. When 352 sera from farmed salmon and 302 sera from farmed trout were tested by both the modified and the original CPE-based assays, overall correlations of 97.72 and 96.03% were, respectively, obtained (96.94% combined). When the modified assay was used to test 188 sera collected from wild salmonids in freshwater river systems in Northern Ireland, no positive results were recorded.
Graham
D A
DA
Veterinary Sciences Division, Department of Agriculture and Rural Development for Northern Ireland, Stormont, Belfast, UK. david.graham@dardni.gov.uk
Jewhurst
V A
VA
Rowley
H M
HM
McLoughlin
M F
MF
Todd
D
D
D000524:Alphavirus / Q000276:immunology / Q000302:isolation & purification*; D018354:Alphavirus Infections / Q000175:diagnosis* / Q000453:epidemiology / Q000276:immunology / Q000662:veterinary*; D000818:Animals; D002460:Cell Line; D005393:Fish Diseases / Q000175:diagnosis* / Q000453:epidemiology / Q000276:immunology / Q000821:virology*; D007124:Immunoenzyme Techniques / Q000379:methods* / Q000662:veterinary*; D009500:Neutralization Tests / Q000379:methods / Q000662:veterinary; D009660:Northern Ireland / Q000453:epidemiology; D017686:Oncorhynchus mykiss / Q000276:immunology / Q000821:virology; D020317:Salmo salar / Q000276:immunology / Q000821:virology; D016036:Seroepidemiologic Studies