Entwicklung und Validierung eines Sandwich ELISA zum Nachweis des rinderspezifischen Knochengewebemarkers Osteocalcin in Fleisch und Knochenmehl
2010
Bernhardt,C.
allemand. Species specific detection and detection of groups of species such as ruminants is requiredaccording to European legislation when dealing with the safe use of animal by-products inanimal nutrition [20]. Various methods are applied to the analysis of feed samples for thepresence of banned processed animal proteins (PAPs) including meat and bone meal(MBM). At present classical microscopy is the official testing method within the EU to detectthe presence of constituents of animal origin in feed for qualitative evaluations of animalfeeds and meat and bone meal (MBM). The method can be applied at two levels. 1.) thedetection of MBM irrespective of the origin and 2.) the detection of MBM from terrestrialanimals in the presence of fishmeal. The results from the DG SANCO 2003 study [18] andfrom the Stratfeed validation study [19] showed that the majority of the participatinglaboratories could not achieve the differentiation power at this taxonomic level. A project ofthe national Referencelaboratories for animal protein in animal feeds (NRL-TP) has the goalto develop an immunological checking method which is based on the ELISA concept. Theprinciple of the immunoassay technique is the interaction between the antibody of the testand the antigen in the sample of the feed. Therefore some anti-bovine osteocalcin polyclonalantibodies of different rabbits were generated, isolated, up-cleaned and finally adapted to theassay. In this elaboration a Sandwich-ELISA concept was used and tested to examinebovine osteocalcin, a new and stable described bone marker protein, to show differentiationbetween specific bone meals but to detect bone meal and PAPs like meat and bone meal ofcattle. In addition the detection of MBM was tested in some legal animal feed, like fishmeal.The Immunoassay was methodically validated to gain basic data for further evaluation.
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Editeur Hochschule für Technik und Wirtschaft Berlin
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