Development of Aptamer Beacons for Antemortem Diagnosis of Chronic Wasting Disease
Abstract
Transmissible spongiform encephalopathies (TSE) such as mad cow disease are transmitted by ingestion of meat contaminated with infective prion protein (PrPsc). TSE of elk and deer termed chronic wasting disease (CWD) has the potential for transmission to human beings. Reliable antemortem diagnostic tests for CWD are necessary for its control. Development of a novel diagnostic probe termed aptamers to detect CWD is proposed. Once selected, the CWD aptamers will be configured as aptamer beacons that can act as molecular switches to turn on a novel and highly sensitive diagnostic technology termed amplifying fluorescing polymer. Objective #1 is to select aptamers for CWD PrPsc using a crossover selection strategy. Initial aptamer selection is being conducted against a tyrosyl- tyrosyl-arginine (YYR) tripeptide exposed in PrPsc but not in PrPc. YYR peptide has been constructed on cellulose, and the selection protocol optimized. Next, selection will target CWD PrPsc that we recently isolated from elk brain. We are currently developing the aptamer selection protocol for PrPsc. The final aptamer selection will crossover and negatively select against recombinant PrPc. Recombinant PrPc will be expressed from a vector constmct containing the elk PrP gene obtained from Dr Katherine O'Rouke at the USDA via a materials transfer agreement.
Document Details
- Document Type
- Technical Report
- Publication Date
- May 01, 2004
- Accession Number
- ADA426544
Entities
People
- Kenneth D. Clinkenbeard
Organizations
- Oklahoma State University–Stillwater