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Hyperefficient PrP Sc amplification of mouse-adapted BSE and scrapie strain by protein misfolding cyclic amplification technique.

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Title: Hyperefficient PrP Sc amplification of mouse-adapted BSE and scrapie strain by protein misfolding cyclic amplification technique.
Authors: Fujihara, Aiko / Atarashi, Ryuichiro / Fuse, Takayuki / Ubagai, Kaori / Nakagaki, Takehiro / Yamaguchi, Naohiro / Ishibashi, Daisuke / Katamine, Shigeru / Nishida, Noriyuki
Issue Date: May-2009
Citation: The FEBS journal, 276(10), pp.2841-2848; 2009
Abstract: Abnormal forms of prion protein (PrP(Sc)) accumulate via structural conversion of normal PrP (PrP(C)) in the progression of transmissible spongiform encephalopathy. Under cell-free conditions, the process can be efficiently replicated using in vitro PrP(Sc) amplification methods, including protein misfolding cyclic amplification. These methods enable ultrasensitive detection of PrP(Sc); however, there remain difficulties in utilizing them in practice. For example, to date, several rounds of protein misfolding cyclic amplification have been necessary to reach maximal sensitivity, which not only take several weeks, but also result in an increased risk of contamination. In this study, we sought to further promote the rate of PrP(Sc) amplification in the protein misfolding cyclic amplification technique using mouse transmissible spongiform encephalopathy models infected with either mouse-adapted bovine spongiform encephalopathy or mouse-adapted scrapie, Chandler strain. Here, we demonstrate that appropriate regulation of sonication dramatically accelerates PrP(Sc) amplification in both strains. In fact, we reached maximum sensitivity, allowing the ultrasensitive detection of < 1 LD(50) of PrP(Sc) in the diluted brain homogenates, after only one or two reaction rounds, and in addition, we detected PrP(Sc) in the plasma of mouse-adapted bovine spongiform encephalopathy-infected mice. We believe that these results will advance the establishment of a fast, ultrasensitive diagnostic test for transmissible spongiform encephalopathies.
Description: The definitive version is available at www.blackwell-synergy.com
Keywords: Prion / Protein misfolding cyclic amplification / Sonication / Transmissible spongiform encephalopathy
URI: http://hdl.handle.net/10069/23318
ISSN: 1742464X
DOI: 10.1111/j.1742-4658.2009.07007.x
PubMed ID: 19459939
Rights: Journal compilation © 2010 Federation of European Biochemical Societies
Type: Journal Article
Text Version: author
Appears in Collections:Articles in academic journal

Citable URI : http://hdl.handle.net/10069/23318

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