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Institute of Infectology (IMED)

National Reference Laboratory for Koi Herpesvirus Disease (KHVD)

The national reference laboratory for KHV disease (KHVD) is located at the Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, and is responsible for coordinating and ensuring the diagnosis of KHVD based on the German regulation of fish diseases and EU legislation. In the EU Directive for aquaculture and in the recommendations of the Office International des Epizooties (OIE) KHVD (KHV disease, KHVD) is listed as a notifiable disease. For differential diagnosis, infections with the Spring Viremia of Carp virus (SVCV), koi sleepy disease (KSD induced by the pathogen carp edema virus (CEV)) and bacterial infections (Aeromonas sp., Pseudomonas sp.) must be considered.

The disease is notifiable after detection of KHV and KHVD in association with the molecular biological detection of the pathogen and its isolation onto cell cultures.

  • Patho- and immunogenesis of Koi Herpesvirus (KHV) disease (KHVD)
  • Diagnosis of KHV-I and KHV with differential diagnosis of SVCV and CEV
  • Carrier fish for KHVD and KSD
  • Investigations on the vaccination of glass eel against viruses with the main point herpesvirus anguillae and birnavirus of eel 
  • Diagnostics of mollusc diseases and oysters (histology, nucleic acid detection)
  1. direct contact for federal and state authorities on KHVD, KSD and SVC diagnostics
  2. Pathogen detection from samples sent to the laboratory
  3. Virus detection in suspected KHVD cases in areas and fish farms which have so far been free from KHV or confirmation of test results of regional laboratories
  4. Application and optimization of the standards, diagnostic procedures, and diagnostic reagents
  5. Testing, standardization and improvement of test procedures for KHV, CEV and SVCV detection
  6. Coordination of the standards and diagnostic procedures used by the veterinary laboratories
  7. Organization of training measures for experts in laboratory diagnosis from veterinary authorities
  8. Organization of comparative national ring tests regarding KHVD
  9. Participation in comparative ring tests for EU Member States organized by the Community reference laboratory
  10. Annual report to the Community reference laboratory on epidemiological and laboratory data on the diagnosis and control of KHVD
  11. Participation in the annual meeting of the representants of the NRL-M of the EU member states
  • KHV antisera
  • Monoclonal antibodies against KHV
  • KHV DNA preparations

The standard procedures used by the NRLs for KHVD and Mollusks diseases for detection:

  • Virus detection (cell culture, immunofluorescence, immunoperoxidase)
  • Genome detection (PCR, RT-PCR, qPCR)
  • Sequence analysis with genotyping
  • NGS with KHV isolates
  • Histology, in-situ hybridization and immunohistochemistry for Mollusks diseases 

Other methods used by the laboratory

  1. Cell culture techniques
  2. Virus cultivation, purification and concentration
  3. Northern, Southern and Western blot
  4. Cloning and sequence analysis
  5. Transfection
  6. Expression of viral genes in pro- and eukaryotic cells
  7. Immunohistochemistry
  8. Flow cytometry and cell sorting

Relevant legal regulations

COMMISSION IMPLEMENTING DECISION (EU) 2015/1554 of 11 September 2015 laying down rules for the application of Directive 2006/88/EC as regards requirements for surveillance and diagnostic methods (notified under document C (2015) 6188)

Council Directive 2006/88/EC on animal health requirements for aquaculture animals and products thereof, and on the prevention and control of certain diseases in aquatic animals (Official Journal of the EU No. L328 p. 14)

Regulation on Notifiable Diseases in the version of the announcement of  Juli 19th 2011 (BGBl. I S. 1404)

Law on the Prevention and Control of Infectious Animal Diseases (Animal Health Law – TierGesG) of May 22nd 2013 (BGBl. I S. 1324 (Nr. 25)

Manual of Diagnostic Tests for Aquatic Animals (2017) des Office International des Epizooties (O.I.E.)

in their latest versions