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, by Commission Implementation Regulation n° 1560/2020 and recently by Commission implementing Regulation (EU) 2022/893. 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.
- Most recent updated and consolidated version (28.06.2022) of Commission Regulation (EC) 152/2009 of 27 January 2009 laying down the methods of sampling and analysis for the official control of feed
- Commission implementing Regulation (EU) 2022/893 of 7 June 2022 amending Annex VI to Regulation (EC) No 152/2009 as regards the methods of analysis for the detection of constituents of terrestrial invertebrates for the official control of feed
- 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, pig and poultry detection by PCR
The calibration kits ERM-AD482 RUMINANT pDNA CALIBRANT, ERM-AD483 PORCINE pDNA CALIBRANT and ERM-AD484k POULTRY pDNA CALIBRANT can be ordered from JRC Geel. They are now Certified European Reference Materials.
For ordering and information, go to the following webpages :
Ruminant calibrant
Porcine calibrant
Poultry calibrant
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
Quote on demand at isales@eurogentec.com - Brilliant II QPCR ROX Master Mix www.agilent.com
“Brilliant II QPCR Low ROX Master Mix” reference number 600806 (Agilent, Santa Clara, CA, USA)
“Brilliant II QPCR High ROX Master Mix” reference number 600805 (Agilent, Santa Clara, CA, USA)
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.
| Title | Version | ||
|---|---|---|---|
| EURL-AP SOP DNA extraction | V1.1 | BINDING | |
| EURL-AP SOP ruminant PCR | V1.3 | BINDING | |
| EURL-AP SOP pig PCR | V1.0 | BINDING | |
| EURL-AP SOP poultry PCR | V1.1 | BINDING | |
| EURL-AP SOP cut-off determination – ruminant | V2.0 | BINDING | |
| EURL-AP SOP cut-off determination – pig | V1.0 | BINDING | |
| EURL-AP SOP cut-off determination – poultry | V1.0 | BINDING | |
| EURL-AP SOP operational schemes | V5.1 | BINDING | |
| EURL-AP SOP observation flowchart | V2.0 | BINDING | |
| EURL-AP SOP slide mounting | V1.0 | BINDING | |
| EURL-AP SOP staining methods | V1.0 | OPTIONAL |