NR AORL
AU Collins,S.J.; Brazier,M.W.; Lewis,V.; Hill,A.F.; Lawson,V.A.; Masters,C.L.
TI Sensitive in vitro detection of protease-resistant prion protein more accurately represents transmissibility in a mouse model of subclinical transmissible spongiform encephalopathy
QU International Conference - Prion diseases: from basic research to intervention concepts - TSE-Forum, 08.10.-10.10.2003, Gasteig, München - Poster session - DG-29
PT Konferenz-Poster
AB The aetiology of sporadic Creutzfeldt-Jakob disease (CJD), which represents approximately 85% of total reported incidences of CJD, has yet to be determined. A range of in vivo models demonstrating extended periods of subclinical infection during which high titres of infectivity are present offer support for covert transmissions as a possible cause of an uncertain number of sporadic CJD cases. Such findings continue to underscore the need for adequately sensitive, readily applicable, screening techniques. Employing quantal dose-titration intracerebral inoculation to generate prolonged subclinical infection in a well characterized model of mouse-adapted human transmissible spongiform encephalopathy (TSE), we assessed the utility of a highly sensitive western immunoblot technique with comparison to conventional methods and to mouse bioassays. By utilising sodium phosphotungstic acid to precipitate PrPsc prior to Western blotting, we were able to detect approximately 250 ID50 units spiked into normal brain homogenate, and this was 50 times more sensitive than conventional techniques. Using this method of Western blotting, we were able to detect PrPsc in the brains of pre-symptomatic mice that had been challenged with low dose inocula and these brains harbored high titres of infectivity, as determined by Tg20 mouse bioasays. We were also able to transmit disease from pre-symptomatic brain homogenates in which we could not detect PrPsc. Comparative studies of this type are important steps in the process of defining optimal screening techniques, especially during the pre-symptomatic phase of TSE infections.
AD S.J. Collins, V. Lewis, C.L. Masters, Australian National Creutzfeldt-Jakob Disease Registry, University of Melbourne, Victoria, Australia 3010; S.J. Collins, M.W. Brazier, V. Lewis, A.F. Hill, V.A. Lawson, C.L. Masters, Department of Pathology, University of Melbourne, Victoria, Australia 3010; A.F. Hill, Department of Biochemistry and Molecular Biology, the University of Melbourne, Victoria, Australia 3010
SP englisch
PO Deutschland