Summary
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 procedures that are used within the MRC Prion Unit 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 brain or peripheral tissues.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Prusiner SB. (1998) Prions. Proc Natl Acad Sci U S A;95:13363–13383.
Collinge J. (2001) Prion diseases of humans and animals: their causes and molecular basis. Annu Rev Neurosci;24:519–550.
Collinge J. (2005) Molecular neurology of prion disease. J Neurol Neurosurg Psychiatry; 76:906–919.
Wadsworth JD, Hill AF, Beck JA, Collinge J. (2003) Molecular and clinical classification of human prion disease. Br Med Bull;66:241–254.
Wadsworth JD, Collinge J. (2007) Update on human prion disease. Biochim Biophys Acta; 1772:598–609.
Weissmann C. (2004) The state of the prion. Nat Rev Microbiol;2:861–871.
Telling GC, Parchi P, DeArmond SJ, et al. (1996) Evidence for the conformation of the pathologic isoform of the prion protein enciphering and propagating prion diversity. Science;274:2079–2082.
Collinge J, Sidle KCL, Meads J, Ironside J, Hill AF. (1996) Molecular analysis of prion strain variation and the aetiology of ‘new variant’ CJD. Nature;383:685–690.
Parchi P, Castellani R, Capellari S, et al. (1996) Molecular basis of phenotypic variability in sporadic Creutzfeldt-Jakob disease. Ann Neurol;39:767–778.
Wadsworth JDF, Hill AF, Joiner S, Jackson GS, Clarke AR, Collinge J. (1999) Strain-specific prion-protein conformation determined by metal ions. Nat Cell Biol;1:55–59.
Parchi P, Giese A, Capellari S, et al. (1999) Classification of sporadic Creutzfeldt-Jakob Disease based on molecular and phenotypic analysis of 300 subjects. Ann Neurol; 46:224–233.
Hill AF, Joiner S, Wadsworth JD, et al. (2003) Molecular classification of sporadic Creutzfeldt-Jakob disease. Brain;126:1333–1346.
Zanusso G, Farinazzo A, Prelli F, et al. (2004) Identification of distinct N-terminal truncated forms of prion protein in different Creutzfeldt-Jakob disease subtypes. J Biol Chem;79: 38936–38942.
Safar JG, Geschwind MD, Deering C, et al. (2005) Diagnosis of human prion disease. Proc Natl Acad Sci U S A;102:3501–3506.
Hill A, Joiner S, Beck J, et al. (2006) Distinct glycoform ratios of protease resistant prion protein associated with PRNP point mutations. Brain;129:676–685.
Lloyd SE, Onwuazor ON, Beck JA, et al. (2001) Identification of multiple quantitative trait loci linked to prion disease incubation period in mice. Proc Natl Acad Sci U S A;98:6279–6283.
Asante EA, Linehan JM, Desbruslais M, et al. (2002) BSE prions propagate as either variant CJD-like or sporadic CJD-like prion strains in transgenic mice expressing human prion protein. EMBO J;21:6358–6366.
Collinge J. (1997) Human prion diseases and bovine spongiform encephalopathy (BSE). Hum Mol Genetics;6:1699–1705.
Collinge J, Palmer.M.S. (1997) Prion Diseases. 1st ed. Oxford: Oxford University Press.
Brown P, Cathala F, Raubertas RF, Gajdusek DC, Castaigne P. (1987) The epidemiology of Creutzfeldt-Jakob disease: conclusion of a 15-year investigation in France and review of the world literature. Neurology;37:895–904.
Collins SJ, Sanchez-Juan P, Masters CL, et al. (2006) Determinants of diagnostic investigation sensitivities across the clinical spectrum of sporadic Creutzfeldt-Jakob disease. Brain;129:2278–2287.
Kovacs GG, Trabattoni G, Hainfellner JA, Ironside JW, Knight RS, Budka H. (2002) Mutations of the prion protein gene phenotypic spectrum. J Neurol; 249:1567–1582.
Mead S. (2006) Prion disease genetics. Eur J Hum Genet;14:273–281.
Brown P, Preece MA, Will RG. (1992) “Friendly fire” in medicine: hormones, homografts, and Creutzfeldt-Jakob disease. Lancet;340:24–27.
Brown P, Preece M, Brandel JP, et al. (2000) Iatrogenic Creutzfeldt-Jakob disease at the millennium. Neurology;55:1075–1081.
Alpers MP. Epidemiology and clinical aspects of kuru. (1987) In: Prusiner SB, McKinley MP, editors. Prions: Novel Infectious Pathogens Causing Scrapie and Creutzfeldt-Jakob Disease. San Diego: Academic Press, 451–465.
Mead S, Stumpf MP, Whitfield J, et al. (2003) Balancing selection at the prion protein gene consistent with prehistoric kurulike epidemics. Science;300:640–643.
Collinge J, Whitfield J, McKintosh E, et al. (2006) Kuru in the 21st century-an acquired human prion disease with very long incubation periods. Lancet;367:2068–2074.
Will RG, Ironside JW, Zeidler M, et al. (1996) A new variant of Creutzfeldt-Jakob disease in the UK. Lancet;347:921–925.
Hill AF, Desbruslais M, Joiner S, et al. (1997) The same prion strain causes vCJD and BSE. Nature;389:448–450.
Bruce ME, Will RG, Ironside JW, et al. (1997) Transmissions to mice indicate that ‘new variant’ CJD is caused by the BSE agent. Nature;389:498–501.
Collinge J. (1999) Variant Creutzfeldt-Jakob disease. Lancet;354:317–323.
Hilton DA, Ghani AC, Conyers L, et al. (2004) Prevalence of lymphoreticular prion protein accumulation in UK tissue samples. J Pathol;203:733–739.
Frosh A, Smith LC, Jackson CJ, et al. (2004) Analysis of 2000 consecutive UK tonsillectomy specimens for disease-related prion protein. Lancet;364:1260–1262.
Hilton DA. (2005) Pathogenesis and prevalence of variant Creutzfeldt-Jakob disease. J Pathol;208:134–141.
Wadsworth JDF, Joiner S, Hill AF, et al. (2001) Tissue distribution of protease resistant prion protein in variant CJD using a highly sensitive immuno-blotting assay. Lancet;358:171–180.
Llewelyn CA, Hewitt PE, Knight RS, et al. (2004) Possible transmission of variant Creutzfeldt-Jakob disease by blood transfusion. Lancet;363:417–421.
Peden AH, Head MW, Ritchie DL, Bell JE, Ironside JW. (2004) Preclinical vCJD after blood transfusion in a PRNP codon 129 heterozygous patient. Lancet; 364:527–529.
Wroe SJ, Pal S, Siddique D, et al. (2006) Clinical presentation and pre-mortem diagnosis of variant Creutzfeldt-Jakob disease associated with blood transfusion: a case report. Lancet;368:2061–2067.
Collinge J, Palmer MS, Dryden AJ. (1991) Genetic predisposition to iatrogenic Creutzfeldt-Jakob disease. Lancet;337:1441–1442.
Palmer MS, Dryden AJ, Hughes JT, Collinge J. (1991) Homozygous prion protein genotype predisposes to sporadic Creutzfeldt-Jakob disease. Nature; 352:340–342.
Windl O, Dempster M, Estibeiro JP, et al. (1996) Genetic basis of Creutzfeldt-Jakob disease in the United Kingdom: a systematic analysis of predisposing mutations and allelic variation in the PRNP gene. Hum Genet;98:259–264.
Lee HS, Brown P, Cervenáková L, et al. (2001) Increased susceptibility to Kuru of carriers of the PRNP 129 methionine/methionine genotype. J Infect Dis;183:192–196.
Wadsworth JD, Asante EA, Desbruslais M, et al. (2004) Human prion protein with valine 129 prevents expression of variant CJD phenotype. Science;306:1793–1796.
Collinge J, Harding AE, Owen F, et al. (1989) Diagnosis of Gerstmann-Sträussler syndrome in familial dementia with prion protein gene analysis. Lancet;2:15–17.
Collinge J, Owen F, Poulter M, et al. (1990) Prion dementia without characteristic pathology. Lancet;336:7–9.
Collinge J, Brown J, Hardy J, et al. (1992) Inherited prion disease with 144 base pair gene insertion: II: clinical and pathological features. Brain;115:687–710.
Mallucci GR, Campbell TA, Dickinson A, et al.(1999) Inherited prion disease with an alanine to valine mutation at codon 117 in the prion protein gene. Brain;122:1823–1837.
Mead S, Poulter M, Beck J, et al. (2006) Inherited prion disease with six octapeptide repeat insertional mutation—molecular analysis of phenotypic heterogeneity. Brain; 129:2297–2317.
World Health Organisation. (2003) WHO manual for surveillance of human transmissible spongiform encephalopathies. http://www.who.int/bloodproducts/TSE-manual2003.pdf
Collinge J, Poulter M, Davis MB, et al. (1991) Presymptomatic detection or exclusion of prion protein gene defects in families with inherited prion diseases. Am J Hum Genet;49:1351–1354.
Budka H, Aguzzi A, Brown P, et al. (1995) Neuropathological diagnostic criteria for Creutzfeldt-Jakob disease (CJD) and other human spongiform encephalopathies (prion diseases). Brain Pathol;5:459–466.
Budka H. Neuropathology of prion diseases. (2003) Br Med Bull;66:121–130.
Hainfellner JA, Brantner-Inthaler S, Cervenáková L, et al. (1995) The original Gerstmann-Sträussler-Scheinker family of Austria: divergent clinicopathological phenotypes but constant PrP genotype. Brain Pathol;5:201–211.
Ironside JW, Head MW. Neuropathology and molecular biology of variant Creutzfeldt-Jakob disease. (2004) Curr Top Microbiol Immunol;284:133–159.
Hill AF, Butterworth RJ, Joiner S, et al. (1999) Investigation of variant Creutzfeldt-Jakob disease and other human prion diseases with tonsil biopsy samples. Lancet;353:183–189.
Glatzel M, Abela E, Maissen M, Aguzzi A. (2003) Extraneural pathologic prion protein in sporadic Creutzfeldt-Jakob disease. N Engl J Med;349:1812–1820.
Hilton DA, Sutak J, Smith ME, et al. (2004) Specificity of lymphoreticular accumulation of prion protein for variant Creutzfeldt-Jakob disease. J Clin Pathol;57:300–302.
Head MW, Ritchie D, Smith N, et al. (2004) Peripheral tissue involvement in sporadic, iatrogenic, and variant Creutzfeldt-Jakob disease: an immunohistochemical, quantitative, and biochemical study. Am J Pathol;164:143–153.
Joiner S, Linehan JM, Brandner S, Wadsworth JD, Collinge J. (2005) High levels of disease related prion protein in the ileum in variant Creutzfeldt-Jakob disease. Gut;54:1506–1508.
Wadsworth JD, Joiner S, Fox K, et al. (2007) Prion infectivity in vCJD rectum. Gut;56:90–94.
Siddique D, Kennedy A, Thomas D, et al. (2005) Tonsil biopsy in the investigation of suspected variant Creutzfeldt-Jakob disease—a cohort study of 50 patients. J Neurol Sci;238:S1–S570.
Advisory Committee on Dangerous Pathogens and the Spongiform Encephalopathy Advisory Committee. (2003) Transmissible spongiform encephalopathy agents: safe working and the prevention of infection. UK Department of Health. http://www.advisorybodies.doh.gov.uk/ acdp/tseguidance/Index.htm
Safar J, Wille H, Itri V, et al. (1998) Eight prion strains have PrPSc molecules with different conformations. Nat Med; 4:1157–1165.
Riesner D. (2003) Biochemistry and structure of PrPC and PrPSc. Br Med Bull;66:21–33.
Parchi P, Capellari S, Chen SG, et al. (1997) Typing prion isoforms. Nature;386:232–233.
Piccardo P, Dlouhy SR, Lievens PMJ, et al. (1998) Phenotypic variability of Gerstmann-Sträussler-Scheinker disease is associated with prion protein heterogeneity. J Neuropathol Exp Neurol;57:979–988.
Parchi P, Chen SG, Brown P, et al. (1998) Different patterns of truncated prion protein fragments correlate with distinct phenotypes in P102L Gerstmann-Sträussler-Scheinker disease. Proc Natl Acad Sci U S A;95:8322–8327.
Furukawa H, Doh-ura K, Kikuchi H, Tateishi J, Iwaki T. (1998) A comparative study of abnormal prion protein isoforms between Gerstmann-Sträussler-Scheinker syndrome and Creutzfeldt-Jakob disease. J Neurol Sci;158:71–75.
Piccardo P, Liepnieks JJ, William A, et al. (2001) Prion proteins with different conformations accumulate in Geustmann-Sträussler-Scheinker disease caused by A117V and F198S mutations. Am J Pathol;158:2201–2207.
Tagliavini F, Lievens PMJ, Tranchant C, et al. (2001) A 7-kDa prion protein (PrP) fragment, an integral component of the PrP region required for infectivity, is the major amyloid protein in Gerstmann-Sträussler-Scheinker disease A117V. J Biol Chem;276:6009–6015.
Wadsworth JD, Joiner S, Linehan JM, et al. (2006) Phenotypic heterogeneity in inherited prion disease (P102L) is associated with differential propagation of protease-resistant wild-type and mutant prion protein. Brain;129:1557–1569.
Acknowledgments
We especially thank all patients and their families for generously consenting to use of human tissues in this research, and the UK neuropathologists who have kindly helped in providing these tissues. We are grateful to R. Young for preparation of the figures. This work was funded by the UK Medical Research Council and the European Commission, and it was performed under approval from the Institute of Neurology/National Hospital for Neurology and Neurosurgery Local Research Ethics Committee.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2008 Humana Press, a part of Springer Science + Business Media, LLC
About this protocol
Cite this protocol
Wadsworth, J.D.F. et al. (2008). Molecular Diagnosis of Human Prion Disease. In: Hill, A.F. (eds) Prion Protein Protocols. Methods in Molecular Biology™, vol 459. Humana Press. https://doi.org/10.1007/978-1-59745-234-2_14
Download citation
DOI: https://doi.org/10.1007/978-1-59745-234-2_14
Publisher Name: Humana Press
Print ISBN: 978-1-58829-897-3
Online ISBN: 978-1-59745-234-2
eBook Packages: Springer Protocols