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@ARTICLE{Bannach:202091,
author = {Bannach, Oliver and Birkmann, Eva and Reinartz, Elke and
Jaeger, Karl-Erich and Langeveld, Jan P. M. and Rohwer,
Robert G. and Gregori, Luisa and Terry, Linda A. and
Willbold, Dieter and Riesner, Detlev},
title = {{D}etection of {P}rion {P}rotein {P}articles in {B}lood
{P}lasma of {S}crapie {I}nfected {S}heep},
journal = {PLoS one},
volume = {7},
number = {5},
issn = {1932-6203},
address = {Lawrence, Kan.},
publisher = {PLoS},
reportid = {FZJ-2015-04380},
pages = {e36620 -},
year = {2012},
abstract = {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.},
cin = {ICS-6},
ddc = {500},
cid = {I:(DE-Juel1)ICS-6-20110106},
pnm = {452 - Structural Biology (POF2-452)},
pid = {G:(DE-HGF)POF2-452},
typ = {PUB:(DE-HGF)16},
UT = {WOS:000305341500089},
pubmed = {pmid:22567169},
doi = {10.1371/journal.pone.0036620},
url = {https://juser.fz-juelich.de/record/202091},
}