%0 Journal Article
%A Birkmann, E.
%A Henke, F.
%A Weinmann, N
%A Dumpitak, C.
%A Groshup, M.
%A Funke, A.
%A Willbold, D.
%A Riesner, D.
%T Counting of single prion particles bound to a capture-antibody surface (surface-FIDA)
%J Veterinary microbiology
%V 123
%@ 0378-1135
%C Amsterdam [u.a.]
%I Elsevier Science
%M PreJuSER-60108
%P 294 - 304
%D 2007
%Z Record converted from VDB: 12.11.2012
%X 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.
%K Animals
%K Blotting, Western: veterinary
%K Cattle
%K Cricetinae
%K Electrophoresis, Polyacrylamide Gel: veterinary
%K Encephalopathy, Bovine Spongiform: diagnosis
%K Endopeptidase K: chemistry
%K Endopeptidase K: metabolism
%K PrPSc Proteins: cerebrospinal fluid
%K PrPSc Proteins: isolation & purification
%K Prion Diseases: diagnosis
%K Prion Diseases: veterinary
%K Prions: isolation & purification
%K Scrapie: diagnosis
%K Sensitivity and Specificity
%K Spectrometry, Fluorescence: methods
%K Spectrometry, Fluorescence: veterinary
%K PrPSc Proteins (NLM Chemicals)
%K Prions (NLM Chemicals)
%K Endopeptidase K (NLM Chemicals)
%K J (WoSType)
%F PUB:(DE-HGF)16
%9 Journal Article
%$ pmid:17499942
%U <Go to ISI:>//WOS:000249306400003
%R 10.1016/j.vetmic.2007.04.001
%U https://juser.fz-juelich.de/record/60108