Detection of ruminant meals in feed by real-time PCR

  • Prado, M. , Berben, G. , Fumière, O. , Van Duijn, G. , Mensinga-Kruize, J. , Reaney, S. , Boix, A. & Von Holst, C. (2007). Detection of ruminant meals in feed by real-time PCR. CRA-W. Proceedings in: FEED SAFETY International Conference 2007: Methods and Challenges, Namur - Belgique, 27-28/11/2007, 70-71.
Type Conference Proceedings
Year of conference 2007
Title Detection of ruminant meals in feed by real-time PCR
Editor CRA-W
Conference name FEED SAFETY International Conference 2007: Methods and Challenges
Conference location Namur - Belgique
Recnumber 1099
Pages 70-71
Label 1099
conference Date 27-28/11/2007
Endnote Keywords Ruminant|beef|feed|animal meals|MBM|mitochondrial/chromosomal DNA|real-time|
Abstract The appearance of bovine spongiform encephalopathy (BSE) led the Commission of the European Communities (EC) to ban the usage of proteins from the same species to feed animals. Enforcing these regulations required analytical methods capable to allow species-specific identification. The lack of such methods led to the introduction of an extended feed ban for all farmed animals by amending Regulation 999/2001, through Commission Regulation 1234/2003. The availability of analytical methods to correctly differentiate animal species is one of the conditions to re-introduce the use of non-ruminant mammalian proteins to feeds for non-ruminant species. So far, classical microscopy is the only official method within the EU to detect the presence of constituents of animal origin, although with a high sensitivity to detect MBM in feeds, the real potential of this method for the discrimination of the material according to the species origin of the materials is questioned. Among alternative techniques, PCR-based methods seem a promising solution for the detection and identification of animal species in feed. Recently an interlaboratory study was organized to check the performance of four available PCR methods in Europe for cattle or ruminant detection. The results of the study indicate that the four methods applied were able to detect 0.1% cattle MBM, either alone or in mixtures with different materials. This result is a clear improvement when compared with results from former interlaboratory studies. The analysis of the results shows that this improvement is related with specific characteristics of the methods which are mainly the small size of amplicons (68 to 142 bp), the use of real-time PCR, which allows the use of smaller targets, and the location of the target, in all cases sequences that can be found in high copy number. All these factors affected positively the detection of MBM in animal feed and can serve as guidance for other laboratories developing methods for MBM detection and animal species identification in feeds.
Author address Berben Gilbert, Quality Department of Agro-food Products, Walloon Agricultural Research Centre (CRA-W), Chaussée de Namur, 24, B-5030 Gembloux,
Caption 1099-berben-2007.pdf
Authors Prado, M., Berben, G., Fumière, O., Van Duijn, G., Mensinga-Kruize, J., Reaney, S., Boix, A., Von Holst, C.