TY  - JOUR
AU  - Birkmann, E.
AU  - Henke, F.
AU  - Weinmann, N
AU  - Dumpitak, C.
AU  - Groshup, M.
AU  - Funke, A.
AU  - Willbold, D.
AU  - Riesner, D.
TI  - Counting of single prion particles bound to a capture-antibody surface (surface-FIDA)
JO  - Veterinary microbiology
VL  - 123
SN  - 0378-1135
CY  - Amsterdam [u.a.]
PB  - Elsevier Science
M1  - PreJuSER-60108
SP  - 294 - 304
PY  - 2007
N1  - Record converted from VDB: 12.11.2012
AB  - 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.
KW  - Animals
KW  - Blotting, Western: veterinary
KW  - Cattle
KW  - Cricetinae
KW  - Electrophoresis, Polyacrylamide Gel: veterinary
KW  - Encephalopathy, Bovine Spongiform: diagnosis
KW  - Endopeptidase K: chemistry
KW  - Endopeptidase K: metabolism
KW  - PrPSc Proteins: cerebrospinal fluid
KW  - PrPSc Proteins: isolation & purification
KW  - Prion Diseases: diagnosis
KW  - Prion Diseases: veterinary
KW  - Prions: isolation & purification
KW  - Scrapie: diagnosis
KW  - Sensitivity and Specificity
KW  - Spectrometry, Fluorescence: methods
KW  - Spectrometry, Fluorescence: veterinary
KW  - PrPSc Proteins (NLM Chemicals)
KW  - Prions (NLM Chemicals)
KW  - Endopeptidase K (NLM Chemicals)
KW  - J (WoSType)
LB  - PUB:(DE-HGF)16
C6  - pmid:17499942
UR  - <Go to ISI:>//WOS:000249306400003
DO  - DOI:10.1016/j.vetmic.2007.04.001
UR  - https://juser.fz-juelich.de/record/60108
ER  -