Molecular Discrimination of Sheep Bovine Spongiform Encephalopathy from Scrapie
Laura Pirisinu, Sergio Migliore, Michele Angelo Di Bari, Elena Esposito, Thierry Baron, Claudia D’Agostino, Luigi De Grossi, Gabriele Vaccari, Umberto Agrimi, and Romolo Nonno
Author affiliations: Author affiliations: Istituto Superiore di Sanità, Rome, Italy (L. Pirisinu, S. Migliore, M.A. Di Bari, E. Esposito, C. D’Agostino, G. Vaccari, U. Agrimi, R. Nonno); Agence Nationale de Sécurité Sanitaire, Lyon, France (T. Baron); Istituto Zooprofilattico Sperimentale delle Regioni Lazio e Toscana, Rome (L. De Grossi)
Figure 1. Representative Western blot showing the differential N-terminal proteinase K cleavage (monoclonal antibodies SAF84 vs. P4) and the susceptibility to denaturation of different transmissible spongiform encephalopathy isolates. Samples are indicated according to Table 2: classical scrapie isolates (Sc1, Sc2, Sc3, Sc4); experimental CH1641 (Ch1); CH1641-like isolates (Ch2, Ch3, Ch4); experimental sheep bovine spongiform encephalopathy by intracerebral transmission (Bs1) and oral transmission (Bs2, Bs3, Bs4, Bs5); natural goat isolate (Bs6). All samples were pretreated (+) or not treated (-) with 3.5 mol/L guanidine hydrochloride for 1 h at 37°C and then diluted to a final concentration of 0.35 mol/L guanidine hydrochloride, before digestion with proteinase K according to the Istituto Superiore di Sanità discriminatory method. Replica blots were probed with SAF84 (top) and P4 (bottom). Molecular weights are indicated on the right. GdnHCl, guanidine hydrochloride.
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