CHARACTERIZATION OF ANTISERA RAISED AGAINST SPECIES-SPECIFIC PEPTIDE SEQUENCES FROM SCRAPIE-ASSOCIATED FIBRIL PROTEIN AND THEIR APPLICATIONFOR POSTMORTEM IMMUNODIAGNOSIS OF SPONGIFORM ENCEPHALOPATHIES

Transmissible spongiform encephalopathies (TSE), such as scrapie or Cr eutzfeldt-Jakob disease (CJD), are fatal neurodegenerative diseases of the central nervous system caused by a yet unidentified virus. They a re accompanied by a brain specific amyloidosis, during which a host co ded protein irreversibly aggregates to form the scrapie-associated fib rils. The diagnosis of TSE relies on histopathological detection of sp ongiform lesions, on electron microscopical detection of fibrils, or o n the immunological detection of SAF protein, which is the most specif ic diagnostic marker. In order to improve the diagnosis of TSE, we dev eloped a protocol for rapid tissue fractionation and enrichment of SAF protein which subsequently allows the specific detection of SAF prote in by western blotting and immunodetection. Using some new antisera ra ised against synthetic peptides with sequences specific for the hamste r, sheep, cattle and human SAF protein, several samples can be diagnos ed for TSE within 24 hours, starting with only 10-100mg of brain tissu e from different species.