Rapid and Quantitative Assay of Amyloid-Seeding Activity in Human Brains Affected with Prion Diseases

• Takatsuki, Hanae

• Satoh, Katsuya

• Sano, Kazunori

• Fuse, Takayuki

• Nakagaki, Takehiro

• Mori, Tsuyoshi

• Ishibashi, Daisuke

• Mihara, Ban

• Takao, Masaki

• Iwasaki, Yasushi

• Yoshida, Mari

• Atarashi, Ryuichiro

• Nishida, Noriyuki

• Gianluigi Zanusso

  editor

抄録

The infectious agents of the transmissible spongiform encephalopathies are composed of amyloidogenic prion protein, PrPSc. Real-time quaking-induced conversion can amplify very small amounts of PrPSc seeds in tissues/body fluids of patients or animals. Using this in vitro PrP-amyloid amplification assay, we quantitated the seeding activity of affected human brains. End-point assay using serially diluted brain homogenates of sporadic Creutzfeldt-Jakob disease patients demonstrated that 50% seeding dose (SD50) is reached approximately 1010/g brain (values varies 108.79-10.63/g). A genetic case (GSS-P102L) yielded a similar level of seeding activity in an autopsy brain sample. The range of PrPSc concentrations in the samples, determined by dot-blot assay, was 0.6-5.4 μg/g brain; therefore, we estimated that 1 SD50 unit was equivalent to 0.06-0.27 fg of PrPSc. The SD50 values of the affected brains dropped more than three orders of magnitude after autoclaving at 121°C. This new method for quantitation of human prion activity provides a new way to reduce the risk of iatrogenic prion transmission.

PLOS ONE, 10(6), e0126930; 2015

収録刊行物

• PLOS ONE

  PLOS ONE 10 (6), e0126930-, 2015-06-12

  Public Library of Science

キーワード

• Multidisciplinary

詳細情報

• CRID: 1050005822275466624
• NII論文ID: 120006986998
• ISSN: 19326203
• DOI: 10.1371/journal.pone.0126930
• HANDLE: 10069/35721
• Web Site: https://nagasaki-u.repo.nii.ac.jp/records/4588
• Web Site: http://dx.plos.org/10.1371/journal.pone.0126930
• 本文言語コード: en
• 資料種別: journal article
• データソース種別:
  • IRDB
  • Crossref
  • CiNii Articles
  • KAKEN