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Methods Mol Biol. 2018;1779:257-263. doi: 10.1007/978-1-4939-7816-8_16.

Amplification and Detection of Minuscule Amounts of Misfolded Prion Protein by Using the Real-Time Quaking-Induced Conversion.

Author information

1
Department of Neurology, University Medicine Goettingen and German Center for Neurodegenerative Diseases (DZNE) - site Göttingen, Göttingen, Germany.
2
Center for Networked Biomedical Research on Neurodegenerative Diseases (CIBERNED), Barcelona, Spain.
3
Department of Neurology, University Medicine Goettingen and German Center for Neurodegenerative Diseases (DZNE) - site Göttingen, Göttingen, Germany. ingazerr@med.uni-goettingen.de.

Abstract

A characteristic feature of transmissible spongiform encephalopathies (TSE) is the progressive accumulation of protein aggregates in the brain in a self-propagation manner. Based on this mechanism, in vitro protein amplification systems (such as real-time quaking-induced conversion (RT-QuIC)) for the detection of misfolded prion protein scrapie (PrPres) in CSF were a major step in pre-mortem diagnosis of human prion diseases. Here, we describe a protocol of the RT-QuIC assay to detect PrPres in CSF of prion disease patients. This methodology depends on prion seeds that induce misfolding and aggregation of a substrate by cycles of incubation and quaking. Besides diagnostics, further applications of the RT-QuIC appear to be promising for discrimination between different PrP subtypes or strains, understanding the mechanism of protein misfolding and pre-screening of anti-prion drugs. The technique can be further developed to be used to study characteristics of misfolded proteins in other "prion like" diseases, such as tauopathies, synucleinopathies, or amyloidopathies.

KEYWORDS:

Cerebrospinal fluid; Creutzfeldt-Jakob disease (CJD); Real-Time Quaking-Induced Conversion (RT-QuIC); Resistant prion protein

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