A novel method for preclinical detection of PrPSc in blood.
In this study, we demonstrate that a moderate amount of protein misfolding cyclic amplification (PMCA) coupled to a novel surround optical fibre immunoassay (SOFIA) detection scheme can be used to detect the disease-associated form of the prion protein (PrP(Sc)) in protease-untreated plasma from preclinical and clinical scrapie sheep, and white-tailed deer with chronic wasting disease, following natural and experimental infection. PrP(Sc), resulting from a conformational change of the normal (cellular) form of prion protein (PrP(C)), is considered central to neuropathogenesis and serves as the only reliable molecular marker for prion disease diagnosis. While the highest levels of PrP(Sc) are present in the central nervous system, the development of a reasonable diagnostic assay requires the use of body fluids that characteristically contain exceedingly low levels of PrP(Sc). PrP(Sc) has been detected in the blood of sick animals by means of PMCA technology. However, repeated cycling over several days, which is necessary for PMCA of blood material, has been reported to result in decreased specificity (false positives). To generate an assay for PrP(Sc) in blood that is both highly sensitive and specific, we have utilized limited serial PMCA (sPMCA) with SOFIA. We did not find any enhancement of sPMCA with the addition of polyadenylic acid nor was it necessary to match the genotypes of the PrP(C) and PrP(Sc) sources for efficient amplification.
Rubenstein
Richard
R
Departments of Neurology and Physiology/Pharmacology, SUNY Downstate Medical Center, 450 Clarkson Avenue, Brooklyn, NY 11203, USA. richard.rubenstein@downstate.edu
Chang
Binggong
B
Gray
Perry
P
Piltch
Martin
M
Bulgin
Marie S
MS
Sorensen-Melson
Sharon
S
Miller
Michael W
MW
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015153:Blotting, Western; D005838:Genotype; D018620:PrPSc Proteins / Q000097:blood* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D017510:Protein Folding; D012608:Scrapie / Q000097:blood* / Q000235:genetics; D012756:Sheep
D000911:Antibodies, Monoclonal; D018620:PrPSc Proteins