Methods for Molecular Diagnosis of Human Prion Disease

Methods Mol Biol. 2017:1658:311-346. doi: 10.1007/978-1-4939-7244-9_22.

Abstract

Human prion diseases are associated with a range of clinical presentations, and they are classified by both clinicopathological syndrome and etiology, with subclassification according to molecular criteria. Here, we describe updated procedures that are currently used within the MRC Prion Unit at UCL to determine a molecular diagnosis of human prion disease. Sequencing of the PRNP open reading frame to establish the presence of pathogenic mutations is described, together with detailed methods for immunoblot or immunohistochemical determination of the presence of abnormal prion protein in the brain or peripheral tissues.

Keywords: Bovine spongiform encephalopathy; Creutzfeldt-Jakob disease; Fatal familial insomnia; Gerstmann-Sträussler-Scheinker disease; Kuru; Prion; Prion disease; Prion protein; Transmissible spongiform encephalopathy; Variant Creutzfeldt-Jakob disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Infective Agents, Local / chemistry
  • Base Sequence
  • Brain / metabolism
  • Brain / pathology
  • Brain Chemistry
  • Electrophoresis, Polyacrylamide Gel / methods
  • Formates / chemistry
  • Gene Expression
  • High-Throughput Nucleotide Sequencing*
  • Humans
  • Immunoblotting / methods
  • Immunohistochemistry / methods*
  • Microtomy / methods
  • Mutation*
  • Open Reading Frames
  • Prion Diseases / diagnosis*
  • Prion Diseases / genetics
  • Prion Diseases / metabolism
  • Prion Diseases / pathology
  • Prion Proteins / genetics*
  • Prion Proteins / metabolism
  • Staining and Labeling / methods*
  • Tissue Embedding / methods

Substances

  • Anti-Infective Agents, Local
  • Formates
  • PRNP protein, human
  • Prion Proteins
  • formic acid