Development of Aptamer Beacons for Antemortem Diagnosis of Chronic Wasting Disease
Annual rept. 1 May 2003-30 Apr 2004
OKLAHOMA STATE UNIV STILLWATER
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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 ORouke at the USDA via a materials transfer agreement.
- Medicine and Medical Research