DAO 90:25-30 (2010)  -  DOI: https://doi.org/10.3354/dao02209

TaqMan® real-time RT-PCR detection of infectious salmon anaemia virus (ISAV) from formalin-fixed paraffin-embedded Atlantic salmon Salmo salar tissues

M. G. Godoy1, F. S. Kibenge2, M. J. Kibenge2, P. Olmos1, L. Ovalle1, A. J. Yañez3, R. Avendaño-Herrera4,*

1Biovac S.A., Puerto Montt, Chile
2Department of Pathology and Microbiology, OIE Reference Laboratory for ISA, Atlantic Veterinary College, University of Prince Edward Island, Charlottetown, C1A 4P3 Prince Edward Island, Canada
3Laboratorio de Enzimología, Instituto de Bioquímica, Facultad de Ciencias, Universidad Austral de Chile Campus Isla Teja, Casilla 567, Valdivia, Chile
4Facultad de Ciencias Biológicas, Departamento de Ciencias Biológicas, Universidad Andres Bello, Viña del Mar, Chile
*Corresponding author. Email: ;

ABSTRACT: The objective of this study was to evaluate the application of a TaqMan® real-time reverse transcriptase PCR (RT-PCR) assay for the detection of infectious salmon anaemia virus (ISAV) in formalin-fixed paraffin-embedded (FFPE) fish tissues from Atlantic salmon Salmo salar with and without clinical signs of infection, and to compare it with histological and immunohistochemical (IHC) techniques. Sixteen fish samples obtained in 2007 and 2008 from 4 different farms in Chile were examined. The real-time RT-PCR allowed the detection of ISAV in FFPE samples from 9 of 16 fish, regardless of the organs analyzed, whereas 4 of the real-time RT-PCR negative fish were positive as indicated by histological examination and 3 of the real-time RT-PCR positive fish were negative as indicated by immunohistochemistry evaluation. The presence of ISAV in RT-PCR positive samples was confirmed by amplicon sequencing. This work constitutes the first report on the use of real-time RT-PCR for the detection of ISAV in FFPE sections. The assay is very useful for the examination of archival wax-embedded tissues, and allows for both prospective and retrospective evaluation of tissue samples for the presence of ISAV. However, the method only confirms the presence of the pathogen and should be used in combination with histopathology, which is a more precise tool. The combination of both techniques would be invaluable for confirmatory diagnosis of infectious salmon anaemia (ISA), which is essential for solving salmon farm problems.


KEY WORDS: Detection · Infectious salmon anaemia virus · Fixed tissue · Real-time RT-PCR


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Cite this article as: Godoy MG, Kibenge FS, Kibenge MJ, Olmos P, Ovalle L, Yañez A, Avendaño-Herrera R (2010) TaqMan® real-time RT-PCR detection of infectious salmon anaemia virus (ISAV) from formalin-fixed paraffin-embedded Atlantic salmon Salmo salar tissues. Dis Aquat Org 90:25-30. https://doi.org/10.3354/dao02209

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