NR AWFX
AU Gough,K.C.; Owen,J.P.; Rees,H.C.; Terry,L.A.; Thorne,L.; Kilpatrick,J.B.; Jackman,R.; Whitelam,G.C.
TI Molecular strain-typing of ovine TSE diseases
QU International Conference - Prion 2006: Strategies, advances and trends towards protection of society - 3.10.-6.10.2006, Torino, Italy, Lingotto Conference Centre - Poster sessions PR-16
PT Konferenz-Poster
AB It is presumed that conformational differences in the PrPsc molecule impart the ability of TSE agents to exist as differing strain types which can be characterised by bioassay. Such three-dimensional structural variations will also dictate their interaction with enzymes and antibodies. At present, methods that detect Proteinase K (PK) resistant forms of PrPsc have been used to differentiate strain-dependent conformers by relative molecular mass, ratios of the different glycosylated forms and differential antibody binding in Western blot analysis and ELISA. However, these methods have not been able to reliably distinguish all variants of scrapie within sheep that have been revealed through bioassay. The present study aims to develop further molecular strain typing assays in order to provide complementary tests capable of differentiating BSE and a range of scrapie strains in sheep of differing genotypes. Rapid means of determining and characterising the different strains of TSE agents is essential for developing control and management strategies of prion diseases in animals and for assessing their risk to man. Here, we describe a novel method, using the thermostable protease thermolysin to digest PrPsc, which reveals PrP banding patterns on Western blots capable of differentiating ovine scrapie field cases and experimental ovine BSE. Such differentiation was apparent in all of the samples analysed, which included brain homogenates from scrapie affected animals of genotypes VRQ/VRQ, ARQ/ARQ, ARQ/VRQ and AHQ/AHQ compared to BSE affected animals with an ARQ/ARQ genotype. The application of such an assay to differentiate experimental scrapie strains is under investigation.
AD K.C. Gough, J.P. Owen, J.B. Kilpatrick: ADAS Rosemaund, Preston Wynne, Hereford. HR1 3PG. UK; H.C. Rees, G.C. Whitelam: Department of Biology, University of Leicester, Leicester. LE1 7RH. UK; L.A. Terry, L. Thorne, R. Jackman: Veterinary Laboratory Agency, Woodham Lane, New Haw, Surrey. KT15 3NB. UK. E-mail: Kevin.Gough@adas.co.uk
SP englisch
PO Italien