Development of novel methods and reagents for surveillence of prion infection

The prion disease process involves a conformational change in the prion protein that in turn can serve as the basis for an early detection, diagnostic, prognostic and screening assay. The Misfolded Protein Diagnostic (MPD) Assay detects infectivity in a crude sample through the use of labeled Pronucleon peptides that undergo conformational change in the presence of the misfolded infectious prion protein. The conformational peptide change is propagated throughout the ensemble of sequence specific Pronucleon peptides in the reaction via a nucleation event resulting in an amplification of signal. Preliminary data supports the simplicity and exquisite sensitivity of this non-immunological approach to infectious prion protein detection. Level of detection has been determined to be near 1 infectious dose for titered murine sCJD plasma samples. During Phase I funding, the MPD Assay has been under optimization with keen focus on defining the positive controls for the assay as well as looking at variability between normal human plasma samples to better define the threshold of detection. With continued funding we propose to define and further optimize the assay reagents and metrics into a platform system that can be reduced to a high-throughput, small volume automated format.