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@ARTICLE{Birkmann:60108,
author = {Birkmann, E. and Henke, F. and Weinmann, N and Dumpitak, C.
and Groshup, M. and Funke, A. and Willbold, D. and Riesner,
D.},
title = {{C}ounting of single prion particles bound to a
capture-antibody surface (surface-{FIDA})},
journal = {Veterinary microbiology},
volume = {123},
issn = {0378-1135},
address = {Amsterdam [u.a.]},
publisher = {Elsevier Science},
reportid = {PreJuSER-60108},
pages = {294 - 304},
year = {2007},
note = {Record converted from VDB: 12.11.2012},
abstract = {Hitherto accredited prion tests use the PK resistance of
PrP(Sc), the pathogenic isoform of the prion protein, as a
marker for the disease. Because of variations in the amount
of disease-related aggregated PrP, which is not
PK-resistant, these prion tests offer only limited
sensitivity. Therefore, a prion detection method that does
not rely on PK digestion would allow for the detection of
both PK-resistant as well as PK-sensitive PrP(Sc).
Furthermore, single particle counting is more sensitive than
methods measuring an integrated signal. Our new test system
is based on dual-colour fluorescence correlation
spectroscopy (FCS). This method quantifies the number of
protein aggregates that have been simultaneously labelled
with two different antibodies using dual-colour fluorescence
intensity distribution analysis (2D-FIDA). This only counts
PrP aggregates, and not PrP monomers. To increase the
sensitivity, PrP(Sc) was concentrated in a two-dimensional
space by immobilizing it so that the antibodies could be
captured on the surface of the slide (surface-FIDA). When
the surface was systematically scanned, even single prion
particles were detected. Using this new technique, the
sensitivity to identify samples from scrapie-infected
hamster as well as BSE-infected cattle can be dramatically
increased in comparison with identification using FIDA in
solution.},
keywords = {Animals / Blotting, Western: veterinary / Cattle /
Cricetinae / Electrophoresis, Polyacrylamide Gel: veterinary
/ Encephalopathy, Bovine Spongiform: diagnosis /
Endopeptidase K: chemistry / Endopeptidase K: metabolism /
PrPSc Proteins: cerebrospinal fluid / PrPSc Proteins:
isolation $\&$ purification / Prion Diseases: diagnosis /
Prion Diseases: veterinary / Prions: isolation $\&$
purification / Scrapie: diagnosis / Sensitivity and
Specificity / Spectrometry, Fluorescence: methods /
Spectrometry, Fluorescence: veterinary / PrPSc Proteins (NLM
Chemicals) / Prions (NLM Chemicals) / Endopeptidase K (NLM
Chemicals) / J (WoSType)},
cin = {INB-2},
ddc = {630},
cid = {I:(DE-Juel1)VDB805},
pnm = {Funktion und Dysfunktion des Nervensystems},
pid = {G:(DE-Juel1)FUEK409},
shelfmark = {Microbiology / Veterinary Sciences},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:17499942},
UT = {WOS:000249306400003},
doi = {10.1016/j.vetmic.2007.04.001},
url = {https://juser.fz-juelich.de/record/60108},
}