Prion diseases, originally called transmissible spongiform encephalopathies, are invariably fatal neurodegenerative diseases that affect humans and animals. The unusual biological and biochemical properties of the infectious agent causing the disease were first recognized in studies of scrapie in sheep, which is the prototypic prion disease. Most cases of prion disease in humans arise sporadically, but some are attributed to PRNP mutations, and still others are caused by zoonotic or iatrogenic transmission. The differential diagnosis of prion disease includes rapidly progressive variants of dementias that are usually chronic and slowly progressive, autoimmune diseases, vascular disease, metabolic diseases, vitamin deficiency, malignancy, and infection. Autopsy is currently the gold standard for the diagnosis and classification of prion diseases and the determination of disease origin. The diagnosis or exclusion of prion disease should be based on the whole clinical picture, including clinical and family history, symptomatology, neurological signs, radiographic findings, and laboratory results.