Ultrasensitive human prion detection in cerebrospinal fluid by real-time quaking-induced conversion

The development of technologies for the in vitro amplification of abnormal conformations of prion protein (PrPSc) has generated the potential for sensitive detection of prions. Here we developed a new PrPSc amplification assay, called real-time quaking-induced conversion (RT-QUIC), which allows the detection of ≥1 fg of PrPSc in diluted Creutzfeldt-Jakob disease (CJD) brain homogenate. Moreover, we assessed the technique first in a series of Japanese subjects and then in a blind study of 30 cerebrospinal fluid specimens from Australia, which achieved greater than 80% sensitivity and 100% specificity. These findings indicate the promising enhanced diagnostic capacity of RT-QUIC in the antemortem evaluation of suspected CJD.

[1]  T. Montine,et al.  A novel human disease with abnormal prion protein sensitive to protease , 2008, Annals of neurology.

[2]  Adriano Aguzzi,et al.  Prions: protein aggregation and infectious diseases. , 2009, Physiological reviews.

[3]  O. Gefeller,et al.  Detection of 14‐3‐3 protein in the cerebrospinal fluid supports the diagnosis of Creutzfeldt‐Jakob disease , 1998, Annals of neurology.

[4]  B. Caughey,et al.  Ultrasensitive detection of scrapie prion protein using seeded conversion of recombinant prion protein , 2007, Nature Methods.

[5]  J. Udgaonkar,et al.  Evidence for stepwise formation of amyloid fibrils by the mouse prion protein. , 2008, Journal of molecular biology.

[6]  David W. Colby,et al.  Prion detection by an amyloid seeding assay , 2007, Proceedings of the National Academy of Sciences.

[7]  K. Abid,et al.  Protein misfolding cyclic amplification for diagnosis and prion propagation studies. , 2006, Methods in enzymology.

[8]  B. Caughey,et al.  Human variant Creutzfeldt-Jakob disease and sheep scrapie PrP(res) detection using seeded conversion of recombinant prion protein. , 2009, Protein engineering, design & selection : PEDS.

[9]  J. Tamada,et al.  Kinetics of insulin aggregation in aqueous solutions upon agitation in the presence of hydrophobic surfaces. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[10]  B. Caughey,et al.  Specific binding of normal prion protein to the scrapie form via a localized domain initiates its conversion to the protease‐resistant state , 1999, The EMBO journal.

[11]  B. Permanne,et al.  Sensitive detection of pathological prion protein by cyclic amplification of protein misfolding , 2001, Nature.

[12]  B. Caughey,et al.  Simplified ultrasensitive prion detection by recombinant PrP conversion with shaking , 2008, Nature Methods.

[13]  P Brown,et al.  Classification of sporadic Creutzfeldt‐Jakob disease based on molecular and phenotypic analysis of 300 subjects , 1999, Annals of neurology.

[14]  W. Surewicz,et al.  Atypical Effect of Salts on the Thermodynamic Stability of Human Prion Protein* , 2003, Journal of Biological Chemistry.

[15]  Stanley B. Prusiner,et al.  Nobel Lecture: Prions , 1998 .

[16]  O. Bocharova,et al.  In vitro conversion of mammalian prion protein into amyloid fibrils displays unusual features. , 2005, Biochemistry.

[17]  D. Brems,et al.  Inverse Relationship of Protein Concentration and Aggregation , 2002, Pharmaceutical Research.

[18]  C. Begue,et al.  Updated clinical diagnostic criteria for sporadic Creutzfeldt-Jakob disease , 2009, Brain : a journal of neurology.

[19]  G. J. Raymond,et al.  N-terminal truncation of the scrapie-associated form of PrP by lysosomal protease(s): implications regarding the site of conversion of PrP to the protease-resistant state , 1991, Journal of virology.

[20]  P. Lansbury,et al.  Cell-free formation of protease-resistant prion protein , 1994, Nature.

[21]  J. Rossier,et al.  Transmission of the BSE Agent to Mice in the Absence of Detectable Abnormal Prion Protein , 1997, Science.

[22]  Manfred Eigen,et al.  Mechanisms of prion protein assembly into amyloid , 2008, Proceedings of the National Academy of Sciences.

[23]  P. Lansbury,et al.  The chemistry of scrapie infection: implications of the 'ice 9' metaphor. , 1995, Chemistry & biology.

[24]  S. Katamine,et al.  Hyperefficient PrPSc amplification of mouse‐adapted BSE and scrapie strain by protein misfolding cyclic amplification technique , 2009, The FEBS journal.

[25]  C. Gibbs,et al.  The 14-3-3 brain protein in cerebrospinal fluid as a marker for transmissible spongiform encephalopathies. , 1996, The New England journal of medicine.

[26]  P. Lansbury,et al.  Aggregates of scrapie-associated prion protein induce the cell-free conversion of protease-sensitive prion protein to the protease-resistant state. , 1995, Chemistry & biology.

[27]  Y. Shiga,et al.  Establishment of a standard 14-3-3 protein assay of cerebrospinal fluid as a diagnostic tool for Creutzfeldt–Jakob disease , 2010, Laboratory Investigation.

[28]  Adam Douglass,et al.  Mechanism of Prion Propagation: Amyloid Growth Occurs by Monomer Addition , 2004, PLoS biology.

[29]  Y. Kuroiwa,et al.  Prospective 10-year surveillance of human prion diseases in Japan. , 2010, Brain : a journal of neurology.

[30]  Leonard A. Smith,et al.  Extreme sensitivity of botulinum neurotoxin domains towards mild agitation. , 2009, Journal of pharmaceutical sciences.

[31]  T. Asada,et al.  Absence of association between codon 129/219 polymorphisms of the prion protein gene and Alzheimer's disease in Japan , 2003, Annals of neurology.

[32]  S. Supattapone,et al.  Ionic Strength and Transition Metals Control PrPSc Protease Resistance and Conversion-inducing Activity* , 2004, Journal of Biological Chemistry.