A Single-Tube HNB-Based Loop-Mediated Isothermal Amplification for the Robust Detection of the Ostreid Herpesvirus 1

Maja A, Zaczek-Moczydlowska, Letitia Mohamed- Smith, Anna Toldra, Chantelle Hooper, Monica Campas, M. Dolors Furones, Tim Bean, Katrina Campbell

Research output: Contribution to journalArticlepeer-review

Abstract

The Ostreid herpesvirus 1 species affects shellfish, contributing significantly to high economic losses during production. To counteract the threat related to mortality, there is a need for the development of novel point-of-care testing (POCT) that can be implemented in aquaculture production to prevent disease outbreaks. In this study, a simple, rapid and specific colorimetric loop-mediated isothermal amplification (LAMP) assay has been developed for the detection of Ostreid herpesvirus 1 (OsHV-1) and its variants infecting Crassostrea gigas (C. gigas). The LAMP assay has been optimized to use hydroxynaphthol blue (HNB) for visual colorimetric distinction of positive and negative templates. The effect of additional Tte UvrD helicase enzyme used in the reaction was also evaluated with an improved reaction time of 10 minutes. Additionally, this study provides a robust workflow for optimization of primers for uncultured viruses using designed target plasmid, when DNA availability is limited.
Original languageEnglish
JournalInternational Journal of Molecular Sciences
DOIs
Publication statusPublished - 9 Sep 2020

Keywords

  • colorimetric
  • Ostreid herpesvirus 1 (OsHV-1
  • loop-mediated isothermal amplification (LAMP)
  • oyster
  • C. gigas
  • early warning detection

Fingerprint

Dive into the research topics of 'A Single-Tube HNB-Based Loop-Mediated Isothermal Amplification for the Robust Detection of the Ostreid Herpesvirus 1'. Together they form a unique fingerprint.

Cite this