NR AXOL
AU Herve,R.; Keevil,B.
TI A Rapid Light Microscopy Technique for Sensitive Detection of Prion Infectivity in Cell Models
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Epidemiology, Risk Assessment and Transmission P04.90
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB Transmissible spongiform encephalopathies (TSEs) are fatal transmissible neurodegenerative diseases observed in various mammal species, including humans. The accumulation of an isoform of the host-encoded prion protein (PrPc) that has become resistant to degradation by proteases is a common feature of TSEs. Although different species possess their own PrP molecule, these are highly conserved and it has become clear that the protease-resistant PrP (PrPsc) is capable of crossing the species barrier, potentially affecting humans. Diagnostic tests rely on immunochemical techniques (Immunohistochemistry, ELISA, Western blotting) using tissues obtained post-mortem. In the mean time, because of the long incubation period, animal models of infectivity are a constraint and pose ethical problems. Therefore, there is a need for a rapid and reliable method allowing the early detection, identification and characterisation of various strains of resistant PrP in animal tissues. In this project, we are assessing an established N2a neuroblastoma prion infectivity model and a Neural Stem Cell (NSC) model developed by our collaborators, which may allow to rapidly assess in vitro the infectivity of various strains of PrPsc. Episcopic Differential Interference Contrast / Epi-Fluorescence (EDIC/EF) microscopy, previously developed in our laboratory, can be used in combination with sensitive fluorescent thiazole dyes to detect proteins particularly rich in beta-sheets. We are using the N2a and NSC models in combination with our detection methods as an alternative to longer and more costly immunochemical protocols. This technique may prove faster, more reliable and more cost effective than current immunochemical methods for rapid confirmation of PrPsc infectivity and assessment of decontamination protocols.
AD R. Hervé, B. Keevil, University of Southampton, Environmental Healthcare Unit, UK
SP englisch
PO Schottland