DAO 80:137-144 (2008)  -  DOI: https://doi.org/10.3354/dao01911

Development of a sensitive and controlled real-time RT-PCR assay for viral haemorrhagic septicaemia virus (VHSV) in marine salmonid aquaculture

Iveta Matejusova*, Paul McKay, Alastair J. A. McBeath, Bertrand Collet, Michael Snow

Fisheries Research Services (FRS) Marine Laboratory, 375 Victoria Road, Aberdeen AB11 9DB, UK

ABSTRACT: A survey was undertaken to determine the potential distribution of viral haemorrhagic septicaemia virus (VHSV) in marine cage-based salmonid farms in Scotland. A rapid, accurate and sensitive quantitative real-time RT-PCR (qRT-PCR) assay was developed, targeting a conserved region of the nucleoprotein (N) gene of the virus. The qRT-PCR assay was shown to be more sensitive than the conventional VHSV RT-PCR. A validation protocol included several different virus isolates as the target and confirmed that the assay could detect all European VHSV genotypes (I, II and III). Both endogenous and exogenous controls were designed to control for integrity of template and distinguish between true VHSV positives and contamination with the positive control material. In total, the universal European VHSV qRT-PCR assay with exogenous positive control was applied to screen 2040 individual Atlantic salmon Salmo salar and 150 individual rainbow trout Oncorhynchus mykiss. No evidence of the presence of VHSV in association with either salmonid species in Scottish marine farms was detected. However, both marine Atlantic salmon and rainbow trout are still considered possible carriers of VHSV, which remains a potential threat to freshwater farming. Therefore, a continued surveillance of these species in marine environment is recommended.


KEY WORDS: Viral haemorrhagic septicaemia virus · VHSV · Atlantic salmon · Rainbow trout · Taqman ·  Real-time PCR · Mariculture


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Cite this article as: Matejusova I, McKay P, McBeath AJA, Collet B, Snow M (2008) Development of a sensitive and controlled real-time RT-PCR assay for viral haemorrhagic septicaemia virus (VHSV) in marine salmonid aquaculture. Dis Aquat Org 80:137-144. https://doi.org/10.3354/dao01911

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