Method of reference for the detection of animal proteins in feed

Annex VI of Commission Regulation n° 152/2009 was revised and amended by Commission Regulation n° 51/2013 and recently amended by Commission Implementation Regulation n° 1560/2020. The new regulation relies on the combination of light microscopy and real time PCR for the detection of animal proteins in feed and on Standard Operating Protocols (SOPs) edited by the EURL-AP for technical implementation.

These SOPs are binding or optional complements to the regulation and are available for download on this page.

  •  Commission Implementation Regulation n°2020/1560 amending Annex VI to Regulation (EC) No 152/2009 laying down the methods of analysis for the determination of constituents of animal origin for the official control of feed
  • Commission Regulation n°051/2013 amending Regulation (EC) No 152/2009 as regards the methods of analysis for the determination of constituents of animal origin for the official control of feed

Calibration kits for ruminant and pig detection by PCR

The calibration kits ERM-AD482 RUMINANT pDNA CALIBRANT and ERM-AD483 PORCINE pDNA CALIBRANT can be ordered from JRC Geel. They are now Certified European Reference Materials.

For ordering and information, go to the following webpages :
https://crm.jrc.ec.europa.eu/p/erm-AD482
https://crm.jrc.ec.europa.eu/p/erm-AD483

List of approved master mixes

Detection of ruminant DNA

As mentioned in the  Commission Regulation (EU) No 51/2013 as well as in the SOP “ Detection of ruminant DNA in feed using real-time PCR”, it is crucial to be careful that the master mix used is fit for the purpose and does not contain bovine serum albumin (BSA) or other reagents susceptible to lead to false results due to presence of animal DNA.

The EURL-AP recommends to use one of the following mastermixes:

  • “Universal mastermix” reference number DMML-D2-D600 (Diagenode, Liège, Belgium) www.diagenode.com
  • “qPCR MasterMix” reference number RT-QP2X-03 (Eurogentec, Seraing, Belgium) www.eurogentec.com   

List of SOPs for download

This list is updated and contains only valid versions.  The binding or optional character of a SOP is indicated.  The original version is labelled V.1.0. Revisions are labelled V.#.#. The period of validity of the different versions are mentioned at the end of the document.

SOPs recently uploaded are tagged NEW for two months after publication date.

TitleVersion
EURL-AP SOP DNA extractionV1.1BINDING
EURL-AP SOP ruminant PCRV1.2BINDING
EURL-AP SOP cut-off determinationV2.0BINDING
EURL-AP SOP operational schemesV4.0BINDINGNEW
EURL-AP SOP observation flowchartV1.0BINDINGNEW
EURL-AP SOP slide mountingV1.0BINDING
EURL-AP SOP staining methodsV1.0OPTIONAL