NR ATJJ
AU Sandmeier,B.; Villmann,C.; Becker,C.M.; Düthorn,T.; Gareis,M.; Pischetsrieder,M.
TI Dot Blot Assay as a novel screening method for the detection of central nervous tissues in meat and meat products
QU International Conference - Prion 2005: Between fundamentals and society's needs - 19.10.-21.10.2005, Congress Center Düsseldorf - Poster Session: Decontamination DEC-16
PT Konferenz-Poster
AB
Bovine spongiform encephalopathy (BSE) and the new variant Creutzfeldt-Jakob disease (vCJD) are most likely transmitted by the consumption of prion-containing tissue of infected animals [1]. Since brain and spinal cord are mainly affected, an important means of prophylaxis of vCJD is, therefore, the elimination of central nervous system (CNS) from human nutrition [2].
In previous works, Myelin Proteolipid protein (PLP) was identified as a suitable target protein for the immunochemical detection of CNS tissues in food. Using a polyclonal anti-PLP antiserum a Western Blot protocol was developed. The Western Blot assay using myelin PLP as marker antigen, allowed a highly specific and very sensitive detection of CNS contaminations in raw meat samples and in processed meat products.
In this study, we established a Dot Blot Assay [3] following sample extraction with organic solvents as a timesaving screening method for the detection of central nervous tissues in food, due to reduced analysis time and simultaneous measurement of up to 80 samples. The assay can be applied without detection system, since a positive reaction is visualized by a colour reaction. No false positive test results were obtained by testing non-CNS tissues like liver, kidney, heart, peripheral nerves and skeletal muscle, whereas high signal intensities were obtained from brain and spinal cord samples. For unprocessed meat samples, thus, the detection limit of the Dot Blot Assay was determined experimentally as 0.03 %. These results could also be verified in a blinded study: CNS concentration intervals of 10 out of 12 meat samples were determined correctly using Dot Blot analysis.
With minor modifications, the Dot Blot method could also be adapted to processed meat samples such as different sausage types. In sausage samples, the detection limit was found to depend on the type of meat product. For cooked bologna type sausages and fermented sausages, the detection limit was below 0.1 %, for cooked liver sausages, it was 0.3 % CNS tissue.
Moreover, the Dot Blot assay was also applicable as a swabbing test for the detection of CNS contaminations on meat and working surfaces.
1. Dormont, D., Prion diseases: pathogenesis and public health concerns. FEBS Lett., 2002. 529: p. 17-21.
2. Heits, G.H., Deutsches Lebensmittelbuch, Leitsätze 2003. 2003, Köln: Bundesanzeiger Verlag.
3. Becker, C.M., W. Hoch, and H. Betz, Sensitive immunoassay shows selective association of peripheral and integral membrane proteins of the inhibitory glycine receptor complex. J Neurochem, 1989. 53(1): p. 124-31.
IN Diesmal haben die Autoren einen Dot Blot Assay für den Nachweis von Myelin-Proteolipid-Protein bzw. ZNS-Gewebe in Nahrungsmitteln entwickelt.
AD Barbara Sandmeier, Monika Pischetsrieder, Department of food chemistry, University Erlangen-Nuremberg, Germany; Carmen Villmann, Cord-Michael Becker, Institute of Biochemistry, University Erlangen-Nuremberg, Germany; Tina Düthorn, Manfred Gareis, Institute of Microbiology and Toxicology, Federal Research Centre for Nutrition and Food, Kulmbach, Germany
SP englisch
PO Deutschland