NR AXQC
AU Koenig,C.; Engemann,C.; Krummrei,U.; Hoffmann,R.
TI Highly Sensitive Detection of Prions in Body Fluids
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Epidemiology, Risk Assessment and Transmission P04.124
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB
Transmissible spongiform encephalopathies (TSEs) are fatal disorders of the central nervous system characterised by the progressive accumulation of an abnormal form of the prion protein (PrP). TSEs include scrapie in sheep, bovine spongiform encephalopathy (BSE) in cattle, and Creutzfeldt-Jakob disease (CJD) in humans. It was shown that BSE can be transmitted by the consumption of meat products from BSE infected cattle to humans causing the new variant of CJD (vCJD). Among humans vCJD can be transferred by blood transfusions and organ transplantations. Due to the slow etiopathology, a general screening of all donors appears obligatory, as the infection risk of a transplant can be unperceived. At present, there are no methods available that allow the routine diagnostic detection of PrPsc in body fluids; this is due to the extremely low concentrations of PrPsc expected in e.g. blood. Therefore, a highly sensitive detection method is necessary.
Such a highly sensitive detection system was established - the priontype(R) TSE combining a peroxidase-based colorimetric ELISA with a detection technique called Immuno-PCR. Using Immuno-PCR, sensitivity of the corresponding classical ELISA was increased 10.000-fold resulting in a detection limit of 10 pg/mL recombinant prion protein even when spiked in body fluids as serum, plasma or cerebrospinal fluid.
The sensitivity of priontype(R) TSE can be further improved by an additionally enrichment of PrP using immunoprecipitation (IP). It was shown that IP can improve the detection limit up to 100-fold. Sensitivity and specificity of both formats were proved by analysing spiked plasma samples (vCJD brain and normal brain) as well as plasma samples from patients with various neurodegenerative diseases and from healthy donors. The results clearly show that the established detection system is an excellent tool for a general screening of blood donations.
AD C. Koenig, C. Engemann, U. Krummrei, Priontype GmbH & Co. KG, Germany; R. Hoffmann, University of Leipzig, Institute of Bioanalytical Chemistry, BBZ, Germany
SP englisch
PO Schottland