NR AOWN
AU McCutcheon,S.; Houston,E.F.; Hunter,N.
TI The use of DELFIA for detection and quantification of PrP in sheep
QU International Conference - Prion diseases: from basic research to intervention concepts - TSE-Forum, 08.10.-10.10.2003, Gasteig, München - Poster session - PG-31
PT Konferenz-Poster
AB
Dissociation enhanced lanthanide fluorometric immunoassay (DELFIA) analysis has been explored as a more sensitive method for detecting PrPsc. This assay relies upon the use of immobilised monoclonal antibodies for antigen capture and a europium (Eu)-labelled detector, allowing the specific detection and quantification of PrP. The assay is based on the conformation-dependent immunoassay (Safar et al., 1998) in which isoforms of PrP are quantified by following antibody binding to the denatured and native forms of the prion protein. The ratio of denatured / native protein can be used as in indicator of PrP conformation. We are currently validating this assay using neuronal and non-neuronal tissue homogenates taken from sheep naturally infected with scrapie. At the onset of clinical signs, animals are killed and a range of tissues taken and ascertained scrapie positive using Western blot techniques. We aim to use DELFIA analysis for quantification of PrPsc in the same tissues. Testing large numbers of infected and uninfected tissues is required to confirm the cut-off ratio that allows accurate discrimination between normal and scrapie-infected tissues. The method will be applied in studies of the peripheral pathogenesis of scrapie in experimentally challenged sheep
(This work has been funded by BBSRC)
AD S. McCutcheon, E.F. Houston, Institute for Animal Health, Compton, UK; N. Hunter, Institute for Animal Health, NPU, Edinburgh, UK
SP englisch
PO Deutschland