Laboratory validation of a lateral flow device for the detection of CyHV-3 antigens in gill swabs

J Virol Methods. 2013 Nov;193(2):679-82. doi: 10.1016/j.jviromet.2013.07.034. Epub 2013 Jul 26.

Abstract

Cyprinid herpesvirus-3 (CyHV-3) induces the highly contagious koi herpesvirus disease (KHVD) and may result in significant economic losses to the ornamental and food-producing carp industry. Suspicion of KHVD is triggered by clinical signs and confirmed using laboratory techniques. The latter are labour- and time-consuming, require specialised equipment and trained personnel. For rapid, on-site detection of CyHV-3, a lateral flow device (LFD) was developed using two monoclonal antibodies directed towards the viral glycoprotein ORF65. The LFD was highly specific with analytical and diagnostic specificities of 100%. Analytical sensitivity ranged between 1.25×10(2) and 2.40×10(4) plaque forming units per ml for isolates originating from geographically distinct regions. In experimentally infected carp, CyHV-3 was detected as early as 4-5 days post infection. Diagnostic sensitivities of 52.6% and 72.2% relative to PCR were recorded, depending on the viral isolate used. When onset of mortality was taken as reference, diagnostic sensitivities increased to 67.0% and 93.3%. The diagnostic sensitivity for freshly found-dead animals was 100%, irrespective of the virus isolate used. Given the high specificity and ease-of-use for on-site detection of CyHV-3, the LFD was regarded fit for purpose as a first-line diagnostic tool for the identification of acute CyHV-3 infections in KHVD affected (koi) carp.

Keywords: Cyprinid herpesvirus-3; Immunochromatographic assay; Koi herpesvirus disease; Lateral flow device.

Publication types

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

MeSH terms

  • Animals
  • Antibodies, Monoclonal
  • Antibodies, Viral
  • Antigens, Viral / analysis*
  • Carps
  • Fish Diseases / diagnosis*
  • Fish Diseases / virology
  • Gills / virology*
  • Herpesviridae / immunology
  • Herpesviridae / isolation & purification*
  • Herpesviridae Infections / diagnosis
  • Herpesviridae Infections / veterinary*
  • Herpesviridae Infections / virology
  • Immunoassay / instrumentation
  • Immunoassay / methods
  • Sensitivity and Specificity
  • Veterinary Medicine / instrumentation*
  • Veterinary Medicine / methods
  • Virology / instrumentation*
  • Virology / methods

Substances

  • Antibodies, Monoclonal
  • Antibodies, Viral
  • Antigens, Viral