NR AOUW
AU Klöhn,P.C.; Stoltze,L.; Flechsig,E.; Enari,M.; Weissmann,C.
TI A Quantitative, Highly Sensitive Cell-Based Infectivity Assay for Mouse Scrapie Prions
QU International Conference - Prion diseases: from basic research to intervention concepts - TSE-Forum, 08.10.-10.10.2003, Gasteig, München - Poster session - DG-15
PT Konferenz-Poster
AB The detection and quantification of prion infectivity is fundamental for the study of transmissible spongiform encephalopathies. Infectivity is usually quantified by bioassays that are based on intracerebral inoculation of mice and are time-consuming, imprecise and costly. We have isolated neuroblastoma N2a sublines highly susceptible to mouse prions, as evidenced by accumulation of infectivity and PrPsc, and developed a quantitative in vitro assay for prion infectivity. In the Scrapie Cell (SC) assay, susceptible N2a cells are exposed to prion-containing samples for 3 days, grown to confluence, split 1:10 three times and the proportion of PrPsc - containing cells is determined using automated counting equipment. In a log/log plot, the dynamic range of these assays is linear over two logs of prion concentrations. The SC assay is about as sensitive as the mouse bioassay, ten times faster, more than two orders of magnitude less expensive and suitable for automation. Scrapie cell assays performed in an end point titration format extend the sensitivity, are quantified in analogy to the mouse bioassay and show that infectivity titers measured in tissue culture and in the mouse are similar.
AD P.-C. Kloehn, L. Stoltze, C. Weissmann, MRC Prion Unit, Department of Neurodegenerative Disease, Institute of Neurology, London, UK; E. Flechsig, Institut für Virologie und Immunbiologie, Universität Würzburg, Germany; M. Enari, National Cancer Center Research Institute Radiobiology Division, Tokyo, Japan
SP englisch
PO Deutschland