NR ABIJ
AU Bieschke,J.G.; Giese,A.; Schulz-Schaeffer,W.J.; Zerr,I.; Poser,S.; Eigen,M.; Kretzschmar,H.A.
TI Ultrasensitive detection of pathological prion protein aggregates by dual-color scanning for intensely fluorescent targets
QU Proceedings of the National Academy of Sciences of the United States of America 2000 May 9; 97(10): 5468-73
IA http://www.pnas.org/cgi/content/full/97/10/5468
PT journal article
AB A definite diagnosis of prion diseases such as Creutzfeldt-Jakob disease (CJD) relies on the detection of pathological prion protein (PrPsc). However, no test for PrPsc in cerebrospinal fluid (CSF) has been available thus far. Based on a setup for confocal dual-color fluorescence correlation spectroscopy, a technique suitable for single molecule detection, we developed a highly sensitive detection method for PrPsc. Pathological prion protein aggregates were labeled by specific antibody probes tagged with fluorescent dyes, resulting in intensely fluorescent targets, which were measured by dual-color fluorescence intensity distribution analysis in a confocal scanning setup. In a diagnostic model system, PrPsc aggregates were detected down to a concentration of 2 pM PrPsc, corresponding to an aggregate concentration of approximately 2 fM, which was more than one order of magnitude more sensitive than Western blot analysis. A PrPsc-specific signal could also be detected in a number of CSF samples from patients with CJD but not in control samples, providing the basis for a rapid and specific test for CJD and other prion diseases. Furthermore, this method could be adapted to the sensitive detection of other disease-associated amyloid aggregates such as in Alzheimer's disease.
MH Animals; Creutzfeldt-Jakob Syndrome/*cerebrospinal fluid/*diagnosis; Cricetinae; Fluorescent Dyes; Humans; Pilot Projects; PrPsc Proteins/*cerebrospinal fluid; Prions/analysis; Reproducibility of Results; Research Support, Non-U.S. Gov't; Scrapie; Sensitivity and Specificity; Spectrometry, Fluorescence/methods
AD Max-Planck-Institute for Biophysical Chemistry, Am Fassberg, 37077 Göttingen, Germany.
SP englisch
PO USA
EA pdf-Datei und HTML-Version