NR AXUW
AU Privat,N.; Laffont-Proust,I.; Faucheux,B.A.; Sazdovitch,V.; Frobert,Y.; Laplanche,J.L.; Grassi,J.; Hauw,J.J.; Haik,S.
TI PrP Immunohistochemistry in Human Prion Diseases: from Antibody Screening to a Standardized fast Immunodiagnosis using Automation
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Pathology and Pathogenesis P03.36
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB Demonstration of pathological prion protein accumulation in the central nervous system is required to establish the diagnosis of transmissible subacute encephalopathies (TSEs). In humans, this is frequently achieved using prion protein (PrP) immunohistochemistry in paraffin-embedded tissue, a technique that requires multiple epitope retrieval and denaturing pre-treatments. In addition to be timeconsuming, this procedure induces tissue alterations that preclude accurate morphological examination. The aim of this study was to simplify the procedure of PrP immunohistochemistry in human tissue, together with increased sensitivity and specificity. We screened a panel of 50 monoclonal antibodies produced using various immunogens (human and ovine recombinant PrP, PrP peptides, denatured scrapieassociated fibrils from 263K-infected Syrian hamsters) and directed against different epitopes along the human PrP sequence. A panel of different forms of genetic, infectious and sporadic TSEs was assessed. The most efficient antibodies were then used in different simplified procedures and checked for their efficiency at 37°. We identified a monoclonal antibody allowing a high specific and fast immunodiagnosis with very limited denaturing pre-treatments. A standardized and reliable fast immunostaining procedure was established using an automated diagnostic system (Nexes, Ventana Medical Systems) and allowed PrP detection in the central nervous system and in tonsil biopsies. It was evaluated in a series of 300 patients with a suspected diagnosis of TSEs and showed high sensitivity and specificity.
AD N. Privat, I. Laffont-Proust, B.A. Faucheux, S. Haik, INSERM, France; V. Sazdovitch, J.-J. Hauw, Salpêtrière Hospital, APHP, France; Y. Frobert, J. Grassi, CEA, France; J.L. Laplanche, Lariboisière Hospital, APHP, France
SP englisch
PO Schottland
EA pdf-Datei und Poster (Postertitel: Human prion diseases: from antibody screening to standardized fast immunodiagnosis using automation)