NR AXQV
AU Langeveld,J.P.M.; Erkens,J.H.F.; Benestad,S.L.; Jacobs,J.G.; van Keulen,L.J.M.; Davidse,A.; Bossers,A.; van Zijderveld,F.G.
TI Small Ruminants with Transmissible Spongiform Encephalopathies Analyzed for Properties of PrPsc and PrPres
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Natural and Experimental Strains P02.40
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB
Due to strain dependent variations disease associated PrP (PrPsc) varies phenotypically. PrPsc is a reliable marker for disease even if its appearance varies depending on epitope exposure, susceptibility to proteases, tissue variations and disease incubation time. There is a clear need to have methods available for efficiently detecting all variants. Therefore, antibodies covering several sites of the ovine PrP sequence are tested to find ways to detect in a single assay as many scrapie variants as possible. This was performed by Western blotting and ELISA on both digested and undigested brain stem homogenates, and under native and denatured conditions. A large set of TSE isolates from sheep including classical scrapie, experimental BSE and atypical cases were subjected to this study. More than 100 negative controls of which 30% with known ages were used to investigate the possible effects of variables like breed, genotype, age, herd and country of origin. Western blotting appeared to be a good tool to detect all types of TSEs although the Nor98 cases are suffering of too stringent conditions of proteinase K, confirming a risk when using such method of detection. A novel ELISA set-up without the use of proteinase K allowed detection of all types of TSEs. In a second stage, the same ELISA approach also could discriminate between the known types of TSE by applying appropriate antibodies, digestion with PK and denaturation. This study offers a unique test set up, where non-digested brain homogenates could be used for detection of TSEs. This observation might lead to applications for general TSE detection.
Acknowledgments: This study was supported by funds from Dutch and Norwegian National Ministries of Agriculture and NeuroPrion EU FP6 framework FOOD-CT-2004506579 NeuroPrion projects SmallRuminantEuroStrains and TSE goat.
AD J.P.M. Langeveld, J.H.F. Erkens, J.G. Jacobs, L.J.M. van Keulen, A. Davidse, A. Bossers, F.G. van Zijderveld, CIDC-Lelystad, WageningenUR, Netherlands; S. Benestad, National Veterinay Institute, Norway
SP englisch
PO Schottland