Rapid capture and detection of ostreid herpesvirus-1 from Pacific oyster Crassostrea gigas and seawater using magnetic beads

PLoS One. 2018 Oct 3;13(10):e0205207. doi: 10.1371/journal.pone.0205207. eCollection 2018.

Abstract

Ostreid herpesvirus-1 (OsHV-1) has been involved in mass mortality episodes of Pacific oysters Crassostrea gigas throughout the world, causing important economic losses to the aquaculture industry. In the present study, magnetic beads (MBs) coated with an anionic polymer were used to capture viable OsHV-1 from two types of naturally infected matrix: oyster homogenate and seawater. Adsorption of the virus on the MBs and characterisation of the MB-virus conjugates was demonstrated by real-time quantitative PCR (qPCR). To study the infective capacity of the captured virus, MB-virus conjugates were injected in the adductor muscle of naïve spat oysters, using oyster homogenate and seawater without MBs as positive controls, and bare MBs and sterile water as negative controls. Mortalities were induced after injection with MB-virus conjugates and in positive controls, whereas no mortalities were recorded in negative controls. Subsequent OsHV-1 DNA and RNA analysis of the oysters by qPCR and reverse transcription qPCR (RT-qPCR), respectively, confirmed that the virus was the responsible for the mortality event and the ability of the MBs to capture viable viral particles. The capture of viable OsHV-1 using MBs is a rapid and easy isolation method and a promising tool, combined with qPCR, to be applied to OsHV-1 detection in aquaculture facilities.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aquaculture / methods*
  • Crassostrea / virology*
  • DNA Viruses / genetics
  • DNA Viruses / isolation & purification*
  • DNA Viruses / pathogenicity
  • DNA, Viral / isolation & purification
  • Immunomagnetic Separation / methods
  • Pacific Ocean
  • Real-Time Polymerase Chain Reaction
  • Seafood / virology*
  • Seawater / virology*
  • Time Factors

Substances

  • DNA, Viral

Supplementary concepts

  • Ostreid herpesvirus 1

Grants and funding

This research has received funding from the European Commission Horizon 2020 (H2020) Framework Programme through grant agreement no 678589 (VIVALDI project). The authors also acknowledge support from CERCA Programme/Generalitat de Catalunya. Anna Toldrà acknowledges IRTA-Universitat Rovira i Virgili-Banco Santander for her PhD grant (2015PMF-PIPF-67).