NR ATJS
AU Jacobs,J.G.; van Keulen,L.J.M.; Erkens,J.; Riepema,K.; van Zijderveld,F.G.; Bossers,A.; Langeveld,J.P.M.
TI Antibodies for discriminatory preclinical diagnosis of ovine scrapie and BSE.
QU International Conference - Prion 2005: Between fundamentals and society's needs - 19.10.-21.10.2005, Congress Center Düsseldorf - Poster Session: Diagnosis DIA-04
PT Konferenz-Poster
AB A practical Western blotting method (Wbl) for rapid surveillance in lymphoid tissues of sheep with scrapie or BSE is presented here, using proteinase K resistant prion protein (PrPres) as a marker for infection and selected site specific antibodies for differentation. Retropharyngeal lymph nodes (RLN) were chosen because of their known early involvement in PrP-deposition, size, accessibility and relative preservation in the head during slaughter. From a range of antibodies with affinities for ovine PrP, the most outstanding - 12B2, P4, 9A2, L42 - were used for further test development. The method yielded a chemical sensitivity approximating or similar to immunohistochemistry (IHC) in preclinical condition of naturally scrapie infected VRQ/VRQ sheep and experimentally BSE infected ARQ/ARQ sheep. For 61 brain positive samples obtained in passive and active surveillance, those with genotypes ARH/VRQ, VRQ/VRQ, and ARQ/VRQ were consistently positive for PrPres in RLN, but unexpectedly not all ARQ/ARQ cases were positive, and not all ARR/VRQ were negative. Though this study is limited in the number of samples analysed (n=101), it anyway points out that preclinical diagnosis of scrapie and BSE in lymph nodes from the head can be obtained by Western blotting much earlier compared to routine diagnosis in brain stem tissue, possibly at 1/3 of the incubation time.
IN Schon nach einem Drittel der Inkubationszeit können die Autoren in retropharyngealen Lymphknoten (Nodus lymphaticus retropharyngeus) per Western blot proteaseresistentes Prionprotein nachweisen und dabei mit einem Satz von Antikörpern sogar zwischen einem bestimmten Scrapie-Typ und BSE-infizierten Schafen unterscheiden. Zur Anreicherung benutzen sie eine Alkoholfällung, weil bei ihnen die Phosphorwolframsäure-Fällung die Proteinase-K-Verdauung störte oder die Banden im Western blot verschob.
AD J.G.Jacobs, L.J.M.van Keulen, J.Erkens, K.Riepema, F.G.van Zijderveld, A.Bossers, J.P.M.Langeveld, CIDC-Lelystad, The Netherlands
SP englisch
PO Deutschland