Characterisation of antisera raised against species-specific peptide sequences from scrapie-associated fibril protein and their application for post-mortem immunodiagnosis of spongiform encephalopathies.
Transmissible spongiform encephalopathies (TSE), such as scrapie or Creutzfeldt-Jakob disease (CJD), are fatal neurodegenerative diseases of the central nervous system caused by a yet unidentified virus. They are accompanied by a brain specific amyloidosis, during which a host coded protein irreversibly aggregates to form the scrapie-associated fibrils. The diagnosis of TSE relies on histopathological detection of spongiform lesions, on electron microscopical detection of fibrils, or on the immunological detection of SAF protein, which is the most specific diagnostic marker. In order to improve the diagnosis of TSE, we developed a protocol for rapid tissue fractionation and enrichment of SAF protein which subsequently allows the specific detection of SAF protein by western blotting and immunodetection. Using some new antisera raised against synthetic peptides with sequences specific for the hamster, sheep, cattle and human SAF protein, several samples can be diagnosed for TSE within 24 hours, starting with only 10-100 mg of brain tissue from different species.
Oberdieck
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U
Robert-Koch-Institut des Bundesgesundheitsamtes, Berlin, Federal Republic of Germany.
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YG
Pocchiari
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Diringer
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H
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology*; D015153:Blotting, Western; D002417:Cattle; D006224:Cricetinae; D006801:Humans; D007106:Immune Sera; D015151:Immunoblotting; D007159:Immunologic Tests; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000276:immunology*; D010446:Peptide Fragments / Q000276:immunology; D016133:Polymerase Chain Reaction; D018622:PrP 27-30 Protein; D017096:Prion Diseases / Q000175:diagnosis* / Q000276:immunology; D012756:Sheep; D013045:Species Specificity
D000914:Antibodies, Viral; D007106:Immune Sera; D009419:Nerve Tissue Proteins; D010446:Peptide Fragments; D018622:PrP 27-30 Protein