NR ATLT
AU Leathers,V.; Velek,K.; Koller,S.; Plourde,L.; Toomik,R.
TI Use of Seprion Capture Technology for the Detection of PrPSC in Scrapie Infected Tissues
QU International Conference - Prion 2005: Between fundamentals and society's needs - 19.10.-21.10.2005, Congress Center Düsseldorf - Poster Session: Diagnosis DIA-57
PT Konferenz-Poster
AB
Misfolding of the normal host prion protein (PrPc) and its subsequent accumulation as a protease-resistant conformer (PrPsc) is a well-documented correlate of transmissible spongiform encephalopathies (TSEs). Proteinase K resistance is commonly leveraged as a method for distinguishing PrPsc from PrPc in most TSE diagnostics on the market. The IDEXX TSE post mortem diagnostics (BSE, CWD and scrapie) do not require proteinase K digestion, but instead use Seprion-capture technology applied to a microtiter plate format. This method utilizes a nonbiological PrPsc-specific ligand that selectively binds PrPsc in the presence of excess PrPc. Captured PrPsc is then detected using an anti-PrP antibody-HRPO conjugate. Sample preparation is limited to homogenization of tissue and the addition of diluent; no other processing is required before applying samples to the assay plate. Assay run time is 3-4 hours, depending on the tissue type under evaluation.
The EU and USDA approved IDEXX Herdchek Bovine Spongiform Encephalopathy Antigen test kit (IDEXX BSE-EIA) was modified with the addition of a conjugate concentrate for use with scrapie tissues (IDEXX BSE-scrapie EIA). All other components in the test were unchanged from the IDEXX BSE-EIA. The specificity of the test on ovine brain tissue was 100% (n> 1500) with the mean of the population greater than 10 standard deviations from the test cut-off. The sensitivity of the test was 100% (n>200) on ovine brain tissue. The IDEXX BSE-scrapie test was also used to successfully detect atypical scrapie and scrapie in lymph node and spleen tissue.
The IDEXX BSE-scrapie test, with its absence of a proteinase K digestion step and minimal handling during sample preparation, provides a sensitive, rapid and easy-to-use method for identifying scrapie positive samples. The simplicity of the method allows straightforward adaptation to automation, making it an ideal tool for screening large numbers of samples. The IDEXX BSE-scrapie EIA has recently completed the European Commission evaluation required for EU approval as a scrapie surveillance method.
IN Die Autoren stellen einen relativ neuen TSE-Test vor, der ohne Antikörper und ohne die problematische Inkubation mit der Proteinase K auskommt, weil ein nicht näher bezeichnetes nicht biologisches Polymer spezifisch das PrPsc aus Gewebehomogenaten heraus fängt. Mit Rinder- und Schafsgehirnen funktioniert der Test sehr sensitiv und spezifisch und hat erwartungsgemäß keine Probleme mit den atypischen Scrapie-Fällen. Mit Lymphgeweben funktioniert der Test bisher deutlich weniger sensitiv und spezifisch. Der neue Test heißt "IDEXX BSE-scrapie EIA" und ist eine Weiterentwicklung des bereits von EU und USDA zugelassenen BSE-Tests "IDEXX BSE-EIA".
AD V.Leathers, K.Velek, S.Koller, L.Plourde, R.Toomik, IDEXX Laboratories, One IDEXX Drive, Westbrook, Maine 04092, USA
SP englisch
PO Deutschland