{
  "url": "https://openlegislation.uk/eu-decision-2015-1554/annex-II/part-1/division-I/division-I.6/division-I.6.3",
  "json_url": "https://openlegislation.uk/eu-decision-2015-1554/annex-II/part-1/division-I/division-I.6/division-I.6.3.json",
  "legislation": "eudn/2015/1554",
  "legislation_title": "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) (Text with EEA relevance)",
  "legislation_url": "https://openlegislation.uk/eu-decision-2015-1554",
  "path": "annex/II/part/1/division/I/division/I.6/division/I.6.3",
  "kind": "division",
  "number": "I.6.3",
  "heading": "Neutralisation",
  "text": "Cells from the collected supernatant shall be removed by centrifugation (2 000 to 4 000 × g) or membrane filtration (0,45 μm) with a low protein binding membrane and the supernatant shall be diluted 1:100 and 1:10 000 in cell culture medium.\nAliquots of a minimum of two supernatant dilutions shall be mixed and incubated for 60 minutes at 15 °C with equal parts of the following reagents separately:\n(a)serum containing group specific antibody against VHSV at a 1:50 (vol:vol) dilution;\n(b)serum containing group specific antibody against IHNV at a 1:50 (vol:vol) dilution;\n(c)pool of antisera against the indigenous serotypes of IPNV at a 1:50 (vol:vol) dilution;\n(d)medium alone (positive control).\nFrom each virus supernatant-serum mixture, at least two cell cultures shall be inoculated with 50 μl each and then incubated at 15 °C. The development of CPE shall be checked as described in point I.5.4.\nVHSV strains and isolates that do not react in neutralisation tests shall be identified by IF or ELISA.\nOther neutralisation tests of proven similar efficacy may be used instead.",
  "extent": "E+W+S+N.I.",
  "status": null,
  "version": "current",
  "annotations": [],
  "outstanding_effects": [],
  "provenance": {
    "source": "legislation.gov.uk",
    "source_url": "https://www.legislation.gov.uk/eudn/2015/1554/annex/II/part/1/division/I/division/I.6/division/I.6.3",
    "retrieved_at": "2026-11-01T15:14:14.330569+00:00"
  },
  "licence": "Open Government Licence v3.0",
  "licence_url": "https://www.nationalarchives.gov.uk/doc/open-government-licence/version/3/",
  "attribution": "Contains public sector information licensed under the Open Government Licence v3.0. Source: legislation.gov.uk, The National Archives."
}