Development and evaluation of a loop-mediated isothermal amplification assay for diagnosis of Cyprinid herpesvirus 2

J Virol Methods. 2013 Dec;194(1-2):206-10. doi: 10.1016/j.jviromet.2013.08.028. Epub 2013 Sep 4.

Abstract

Goldfish Haematopoietic Necrosis caused by Cyprinid herpesvirus 2 (CyHV-2) is a severe fish disease with high level of mortality. This is the first study on detection of this disease by loop-mediated isothermal amplification (LAMP). A set of six primers targeting terminase gene (accession no. EU349285.1) was determined after a serial of tests. Detection limit was 1.09 × 10(-4)μg/μL, which was superior to conventional PCR and real-time PCR. No cross reaction with 28 other viruses or bacteria commonly found in fish was observed. The application of commercial kit and instrument for the LAMP assay could reduce the risk of cross contamination, which is suitable for detection of infection under field conditions.

Keywords: CyHV-2; Diagnosis; GFHNV; LAMP.

Publication types

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

MeSH terms

  • Animals
  • DNA Primers / genetics
  • DNA, Viral / genetics
  • DNA, Viral / isolation & purification*
  • Fish Diseases / diagnosis*
  • Fish Diseases / virology*
  • Goldfish
  • Herpesviridae / genetics
  • Herpesviridae / isolation & purification*
  • Herpesviridae Infections / diagnosis
  • Herpesviridae Infections / veterinary*
  • Herpesviridae Infections / virology
  • Nucleic Acid Amplification Techniques / methods*
  • Sensitivity and Specificity

Substances

  • DNA Primers
  • DNA, Viral