PAD-Beads enrichment enhances detection of PrPSc using real-time quaking-induced conversion.

Objective: Scrapie is a transmissible spongiform encephalopathy (TSE) that naturally occurs in sheep and goats. This fatal neurodegenerative disease results from misfolding of the normal cellular prion protein (PrP^C) to a pathogenic prion protein form (PrP^Sc). This pathogenic form, PrP^Sc, accumulates in the brain and lymphoid tissues. The presence of PrP^Sc can be detected by an in vitro conversion assay known as real-time quaking induced conversion (RT-QuIC). RT-QuIC has been used to detect PrP^Sc in a variety of biological tissues from brains to fluids. While this technique is both rapid and sensitive, enhancing the detection of prions would be valuable in the diagnostic laboratories. Results: In this study, we assessed whether PrP^Sc detection sensitivity of RT-QuIC can be increased by enriching PrP^Sc in scrapie tissue homogenates using commercially available aggregated protein binding ligands coated magnetic beads (PAD-Beads). Coupling of RT-QuIC to PAD-Beads based cleanup allowed detection of PrP^Sc rapidly and without dilution of scrapie sheep brain homogenates prior to RT-QuIC. The PAD-Beads sample pretreatment step prior to RT-QuIC is a useful enhancement in the diagnosis of TSEs. [ABSTRACT FROM AUTHOR]