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Diseases of Aquatic Organisms

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DAO 73:23-31 (2006)  -  doi:10.3354/dao073023

Genotyping of marine viral haemorrhagic septicaemia virus isolated from the Flemish Cap by nucleotide sequence analysis and restriction fragment length polymorphism patterns

C. López-Vázquez1, R. S. Raynard2, N. Bain2, M. Snow2, I. Bandín1, C. P. Dopazo1,*

1Unidad de Ictiopatología, Instituto de Acuicultura, Departamento de Microbiología y Parasitología, Universidad de Santiago de Compostela, 15782 Santiago de Compostela, Spain
2Fisheries Research Services Marine Laboratory, PO Box 101, 375 Victoria Road, Aberdeen AB11 9DB, UK
*Corresponding author. Email:

ABSTRACT: A total of 14 viral haemorrhagic septicaemia virus (VHSV) isolates obtained from Greenland halibut Reinhardtius hippoglossoides caught at the Flemish Cap, a fishing ground in the North Atlantic Ocean near Newfoundland, were characterised using restriction fragment length polymorphism (RFLP) and nucleotide sequence analysis. RFLP analysis was performed on a 1259 bp fragment of the glycoprotein (G) gene, and a 305 nucleotide region within the nucleoprotein (N) gene was used for sequence analysis. Representative strains of the 4 established genotypes were employed for comparative purposes. Sequencing analysis indicated that the Flemish cap isolates grouped in Genotype 3, which also includes isolates from wild fish caught in the North Sea and coastal waters of the UK and Ireland, isolates derived from outbreaks of VHS in turbot farms in the British Isles, and an isolate from European eel Anguilla anguilla caught in northern France. Characterisation using RFLPs resulted in the development of a simple and reliable method of typing VHSV at the genotype level using a 2-step restriction analysis (2-SRA) assay.

KEY WORDS: Viral haemorrhagic septicaemia virus · VHSV · Wild fish · Genotyping · Restriction fragment length polymorphism · RFLP · Sequencing analysis · Phylogenetics

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