Document Detail


Detection of prion protein particles in blood plasma of scrapie infected sheep.
MedLine Citation:
PMID:  22567169     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Prion diseases are transmissible neurodegenerative diseases affecting humans and animals. The agent of the disease is the prion consisting mainly, if not solely, of a misfolded and aggregated isoform of the host-encoded prion protein (PrP). Transmission of prions can occur naturally but also accidentally, e.g. by blood transfusion, which has raised serious concerns about blood product safety and emphasized the need for a reliable diagnostic test. In this report we present a method based on surface-FIDA (fluorescence intensity distribution analysis), that exploits the high state of molecular aggregation of PrP as an unequivocal diagnostic marker of the disease, and show that it can detect infection in blood. To prepare PrP aggregates from blood plasma we introduced a detergent and lipase treatment to separate PrP from blood lipophilic components. Prion protein aggregates were subsequently precipitated by phosphotungstic acid, immobilized on a glass surface by covalently bound capture antibodies, and finally labeled with fluorescent antibody probes. Individual PrP aggregates were visualized by laser scanning microscopy where signal intensity was proportional to aggregate size. After signal processing to remove the background from low fluorescence particles, fluorescence intensities of all remaining PrP particles were summed. We detected PrP aggregates in plasma samples from six out of ten scrapie-positive sheep with no false positives from uninfected sheep. Applying simultaneous intensity and size discrimination, ten out of ten samples from scrapie sheep could be differentiated from uninfected sheep. The implications for ante mortem diagnosis of prion diseases are discussed.
Authors:
Oliver Bannach; Eva Birkmann; Elke Reinartz; Karl-Erich Jaeger; Jan P M Langeveld; Robert G Rohwer; Luisa Gregori; Linda A Terry; Dieter Willbold; Detlev Riesner
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2012-05-02
Journal Detail:
Title:  PloS one     Volume:  7     ISSN:  1932-6203     ISO Abbreviation:  PLoS ONE     Publication Date:  2012  
Date Detail:
Created Date:  2012-05-08     Completed Date:  2012-09-17     Revised Date:  2013-06-25    
Medline Journal Info:
Nlm Unique ID:  101285081     Medline TA:  PLoS One     Country:  United States    
Other Details:
Languages:  eng     Pagination:  e36620     Citation Subset:  IM    
Affiliation:
Institute of Physical Biology, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany. bannacho@hhu.de
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MeSH Terms
Descriptor/Qualifier:
Animals
Prions / blood*
Scrapie / blood*
Sheep
Grant Support
ID/Acronym/Agency:
N01-NS-0-2327/NS/NINDS NIH HHS
Chemical
Reg. No./Substance:
0/Prions
Comments/Corrections

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