NR ATLH
AU Lachmann,I.; Grundmann,C.; Osman,A.A.; Lücker,E.
TI Quantitative detection of PrPres in meat products
QU International Conference - Prion 2005: Between fundamentals and society's needs - 19.10.-21.10.2005, Congress Center Düsseldorf - Poster Session: Diagnosis DIA-45
PT Konferenz-Poster
AB
Following the emergence of a new variant of Creutzfeldt-Jakob disease, methods for the detection of tissues of the central nervous system (CNS) as specified risk material in meat products were developed and validated. These methods, however, are not able to detect any actual health hazard as posed by the presence of abnormal prion protein (PrPres). Thus, methods for the detection of PrPres should be evaluated for their potential applicability to the matrix of heat treated meat products. For that purpose it was necessary to develop a micro-technology suitable for the manufacture of meat products in high security laboratories. In previous studies it was possible to produce such standard micro-sausages containing well defined and homogeneously distributed amounts of bovine BSE-positive brain as a basis for validation studies. Bovine BSE-positive CNS was detectable down to a content of 0.25% in heat treated meat products using one of the conventional BSE rapid tests (Biorad, Germany) without major adaptation of the analytical procedure.
In the present study we developed an ultra-sensitive two-site (sandwich) ELISA system using two different monoclonal anti-prion antibodies and Streptavidin-Peroxidase. The detection limit of this system in combination with the purification kit of the newly approved BetaPrion BSE Test Kit is 10-50 pg/ml of recombinant prion protein. The results obtained with this ELISA system for the matrix of micro-sausages are very promising. In particular, sensitivity was distinctly increased. The cut-off for negative sausage standards was 0.084 OD (99.9% statistical security, n = 45) and the limit of detectable BSE-positive CNS was decreased to 0.08% using standard micro sausages containing 0 to 1.00% CNS (r2=0.96).
IN Die Autoren berichten, mit einem modifizierten ELISA-Verfahren Kontaminationen 0,08% BSE-positivem Hirngewebe in einer Wurst nachweisen zu können.
AD I.Lachmann, A.A.Osman, AJ ROBOSCREEN Gesellschaft für molekulare Biotechnologie mbH, Germany; C.Grundmann, E.Lücker, Institute of Food Hygiene, University of Leipzig, Germany
SP englisch
PO Deutschland