RT Journal Article SR Electronic(1) A1 Rubenstein, Richard A1 Chang, Binggong A1 Gray, Perry A1 Piltch, Martin A1 Bulgin, Marie S. A1 Sorensen-Melson, Sharon A1 Miller, Michael W.YR 2010 T1 A novel method for preclinical detection of PrPSc in blood JF Journal of General Virology, VO 91 IS 7 SP 1883 OP 1892 DO https://doi.org/10.1099/vir.0.020164-0 PB Microbiology Society, SN 1465-2099, AB 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 (PrPSc) in protease-untreated plasma from preclinical and clinical scrapie sheep, and white-tailed deer with chronic wasting disease, following natural and experimental infection. PrPSc, resulting from a conformational change of the normal (cellular) form of prion protein (PrPC), is considered central to neuropathogenesis and serves as the only reliable molecular marker for prion disease diagnosis. While the highest levels of PrPSc 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 PrPSc. PrPSc 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 PrPSc 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 PrPC and PrPSc sources for efficient amplification., UL https://www.microbiologyresearch.org/content/journal/jgv/10.1099/vir.0.020164-0