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Biological Chemistry

Editor-in-Chief: Brüne, Bernhard

Editorial Board Member: Buchner, Johannes / Ludwig, Stephan / Sies, Helmut / Turk, Boris / Wittinghofer, Alfred

12 Issues per year


IMPACT FACTOR increased in 2014: 3.268
Rank 106 out of 289 in category Biochemistry & Molecular Biology in the 2014 Thomson Reuters Journal Citation Report/Science Edition

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Detection of prion particles in samples of BSE and scrapie by fluorescence correlation spectroscopy without proteinase K digestion

Eva Birkmann1 / Oliver Schäfer2 / Nicole Weinmann3 / Christian Dumpitak4 / Michael Beekes5 / Roy Jackman6 / Leigh Thorne7 / Detlev Riesner8

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Corresponding author

Citation Information: Biological Chemistry. Volume 387, Issue 1, Pages 95–102, ISSN (Online) 1437-4315, ISSN (Print) 1431-6730, DOI: 10.1515/BC.2006.013, January 2006

Publication History

Received:
June 30, 2005
Accepted:
September 28, 2005
Published Online:
2006-01-13

Abstract

A characteristic feature of prion diseases such as bovine spongiform encephalopathy (BSE) is the accumulation of a pathological isoform of the host-encoded prion protein, PrP. In contrast to its cellular isoform PrPC, the pathological isoform PrPSc forms insoluble aggregates. All commercial BSE tests currently used for routine testing are based on the proteinase K (PK) resistance of PrP, but not all pathological PrP is PK-resistant. In the present study, single prion particles were counted by fluorescence correlation spectroscopy (FCS). The property of PK resistance is not required, i.e., both the PK-resistant and the PK-sensitive parts of the prion particles are detectable. PrP aggregates were prepared from the brains of BSE-infected cattle, as well as from scrapie-infected hamsters, by the NaPTA precipitation method without PK digestion. They were labeled using two different PrP-specific antibodies for FCS measurements in the dual-color mode (2D-FIDA). Within the limited number of samples tested, BSE-infected cattle and scrapie-infected hamsters in the clinical stage of the disease could be distinguished with 100% specificity from a control group. Thus, a diagnostic tool for BSE detection with complete avoidance of PK treatment is presented, which should have particular advantages for testing animals in the preclinical stage.

Keywords: BSE diagnosis; 2D-FIDA; fluorescence correlation spectroscopy; prion; proteinase; scrapie; single particle detection

Citing Articles

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[11]
Giannantonio Panza, Christian Dumpitak, and Eva Birkmann
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[12]
Kensuke Sasaki, Haruhiko Minaki, and Toru Iwaki
The Journal of Pathology, 2009, Volume 219, Number 1, Page 123
[13]
D. W. Colby, Q. Zhang, S. Wang, D. Groth, G. Legname, D. Riesner, and S. B. Prusiner
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[14]
J. Stohr, N. Weinmann, H. Wille, T. Kaimann, L. Nagel-Steger, E. Birkmann, G. Panza, S. B. Prusiner, M. Eigen, and D. Riesner
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