Real-time RT-qPCR assay to detect sequences in the Piscine orthoreovirus-1 genome segment S1 associated with heart and skeletal muscle inflammation in Atlantic salmon

J Fish Dis. 2020 Aug;43(8):955-962. doi: 10.1111/jfd.13205. Epub 2020 Jul 1.

Abstract

During the last decade, Piscine orthoreovirus was identified as the main causative agent of heart and skeletal muscle inflammation (HSMI) in Atlantic Salmon, Norway. A recent study showed that PRV-1 sequences from salmonid collected in North Atlantic Pacific Coast (NAPC) grouped separately from the Norwegian sequences found in Atlantic Salmon diagnosed with HSMI. Currently, the routine assay used to screen for PRV-1 in NAPC water and worldwide cannot differentiate between the two groups of PRV-1. Therefore, this study aimed at developing a real-time polymerase chain reaction (RT-qPCR) assay to target the PRV-1 genome segments specific for variants associated with HSMI. The assay was optimized and tested against 71 tissue samples collected from different regions including Norway, Chile and both coast of Canada and different hosts farmed Atlantic Salmon, wild Coho Salmon and escaped Atlantic Salmon collected in British Columbia, West Coast of Canada. This assay has the potential to be used for screening salmonids and non-salmonids that may carry PRV-1 potentially causing HSMI.

Keywords: genotype; heart and skeletal muscle inflammation; molecular diagnostics; piscine reovirus; quantitative real-time PCR.

MeSH terms

  • Animals
  • Canada
  • Cardiomyopathies / immunology
  • Cardiomyopathies / veterinary*
  • Chile
  • Fish Diseases / immunology
  • Fish Diseases / virology*
  • Inflammation / immunology
  • Inflammation / veterinary*
  • Inflammation / virology
  • Muscle, Skeletal / immunology
  • Muscular Diseases / immunology
  • Muscular Diseases / veterinary*
  • Myocardium / immunology
  • Norway
  • Orthoreovirus / genetics*
  • Real-Time Polymerase Chain Reaction / veterinary
  • Reoviridae Infections / immunology
  • Reoviridae Infections / veterinary*
  • Reoviridae Infections / virology
  • Reverse Transcriptase Polymerase Chain Reaction / veterinary
  • Salmo salar*

Supplementary concepts

  • Piscine orthoreovirus