Detection and discrimination of PrPSc by multi-spectral ultraviolet fluorescence.
Prion diseases are progressive degenerative disorders of the central nervous system. The transmissibility and fatal nature of these diseases necessitate their rapid and accurate diagnosis. The hallmark of these diseases is the accumulation of PrPSc, a protease-resistant form of a host-coded glycoprotein. We have been evaluating the use of multi-spectral ultraviolet fluorescent spectroscopy as a means of detecting and distinguishing between different forms of PrPSc. Spectroscopic measurements of fluorescence from untreated and proteinase K (PK)-treated PrPSc, purified from 263K scrapie strain-infected hamster brains and ME7 scrapie strain-infected mouse brains, were performed. Spectra of untreated and PK-treated PrPSc samples for 263K and ME7 appeared qualitatively different. The identification and discrimination of PrPSc were possible based on these spectral signatures, calculations of their fluorescence cross sections, and determination of the orthogonal differences. This technique has the potential not only for the sensitive, specific, and direct detection of PrPSc, but also for the ability to distinguish between different forms of the prion protein.
Rubenstein
R
R
New York State Institute for Basic Research in Developmental Disabilities, Staten Island, New York 10312, USA.
Gray
P C
PC
Wehlburg
C M
CM
Wagner
J S
JS
Tisone
G C
GC
D000818:Animals; D001921:Brain / Q000378:metabolism; D006224:Cricetinae; D019286:Endopeptidase K; D005069:Evaluation Studies as Topic; D051379:Mice; D018620:PrPSc Proteins / Q000032:analysis* / Q000302:isolation & purification; D012608:Scrapie / Q000175:diagnosis / Q000378:metabolism; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods* / Q000706:statistics & numerical data; D013056:Spectrophotometry, Ultraviolet / Q000379:methods* / Q000706:statistics & numerical data
D018620:PrPSc Proteins; D019286:Endopeptidase K