title: Enrichment of PrPSc in formalin-fixed, paraffin-embedded tissues prior to analysis by Western blot.
abstract: Diagnosis of prion disease is primarily through immunodetection of the infectious agent. Typically, 2 distinct procedures are recommended for a definitive diagnosis, with immunohistochemistry and Western blot providing the most information as to the specific isolate in question. In the past, these approaches required formalin-fixed, paraffin-embedded tissue and fresh or frozen tissue, respectively; however, methods have been developed that allow for use of fixed tissue for Western blot. The present study describes a method of enriching PrP(Sc) in formalin-fixed, paraffin-embedded tissues prior to Western blot analysis for the detection of PrP(Sc). With this modified procedure, 5 times the previously reported sample size may be used for analysis, greatly enhancing the sensitivity of this procedure.
authors: Virus and Prion Research Unit, National Animal Disease Center, ARS, USDA, 1920 Dayton Avenue, PO Box 70, Ames, IA 50010, USA. Eric.Nicholson@ars.usda.gov
authors: Eric M
authors: EM
authors: Nicholson
mesh_terms: D000818:Animals; D015153:Blotting, Western / Q000379:methods / Q000662:veterinary*; D001933:Brain Stem / Q000473:pathology; D005557:Formaldehyde; D016612:Paraffin Embedding / Q000379:methods / Q000662:veterinary*; D018620:PrPSc Proteins / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D012756:Sheep; D016707:Tissue Fixation / Q000379:methods / Q000662:veterinary*
chemical_list: D018620:PrPSc Proteins; D005557:Formaldehyde