NR AOQL
AU Bieschke,J.G.; Weber,P.; Giese,A.; Bertsch,U.; Kretzschmar,H.A.
TI Single molecule detection and characterization of PrPsc - towards a blood test for prion disease
QU International Conference - Prion diseases: from basic research to intervention concepts - TSE-Forum, 08.10.-10.10.2003, Gasteig, München - Poster session - DG-57
PT Konferenz-Poster
AB A definite diagnosis of prion diseases such as Creutzfeldt-Jakob disease (CJD) relies on the detection of pathological prion protein (PrPsc). Based on a setup for confocal dual-color fluorescence correlation spectroscopy (FCS), we developed a highly sensitive detection method for PrPsc. Pathological prion protein aggregates were labeled by specific antibody probes tagged with fluorescent dyes, resulting in intensely fluorescent targets which were measured by dual-color SIFT (scanning for intensly fluorescent targets) setup. However, as the aim of our studies is to detect PrPsc in blood and blood fractions, further improvements in detection sensitivity become necessary. Therefore PrPsc detection has to be coupled with concentration or amplification procedures. We optimised several purification procedures for PrPsc for the use in SIFT detection, which allowed a 100-fold more sensitive detection of PrPsc than Western Blot analysis. A number of of antibody probes were evaluated in respect to detection sensitivity and the potential to differentiate between prion strains. Combination of prion amplification with SIFT-detection promises to provide the sensitivity necessary for the detection of PrPsc in blood.
AD Jan G. Bieschke, Petra Weber, Armin Giese, Uwe Bertsch, Hans A. Kretzschmar, Institute for Neuropathology, LMU Munich, Germany
SP englisch
PO Deutschland