Detection of the disease-associated isoform of the prion protein in formalin-fixed tissues by Western blot.
Clinical signs of prion disease are not specific and include a variety of differential diagnoses. Serological tests and nucleic acid-based detection methods are not applicable to prion-disease-agent detection because of the unusual nature of the infectious agent. Prion-disease diagnosis is primarily conducted by means of immunodetection of the infectious agent, typically by at least 2 distinct procedures with immunohistochemistry and Western blot being the most informative. These approaches differ in the need for formalin-fixed and frozen or fresh tissue respectively. This work describes a method for the detection of the disease-associated isoform of the prion protein by Western blot using formalin-fixed tissues. The approach requires only minimal modification of existing Western-blot procedures and could readily be incorporated into existing detection schemes for confirmatory purposes when fresh or frozen tissues are unavailable.
Nicholson
Eric M
EM
Virus and Prion Diseases of Livestock Research Unit, National Animal Disease Center, USDA, Agricultural Research Service, Ames, IA 50010, USA. eric.nicholson@ars.usda.gov
Kunkle
Robert A
RA
Hamir
Amir N
AN
Lebepe-Mazur
Semakaleng
S
Orcutt
Dennis
D
D000818:Animals; D015153:Blotting, Western / Q000662:veterinary*; D001933:Brain Stem / Q000737:chemistry; D005557:Formaldehyde; D018620:PrPSc Proteins / Q000032:analysis*; D020033:Protein Isoforms; D012608:Scrapie / Q000175:diagnosis; D012756:Sheep; D013997:Time Factors; D016707:Tissue Fixation / Q000662:veterinary*