Development of Aptamer Beacons for Antemortem Diagnosis of Chronic Wasting Disease
Annual rept. 1 May 2004-30 Apr 2005
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. A 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. Initial aptamer selection has been conducted against a tyrosyl-tyrosyl-arginine YYR tripeptide thought to be exposed in PrPSc but not in PrPC. Next, selection will target CWD PrPSc using a novel electrodialysis SELEX technique being developed in our laboratory. The final aptamer selection will crossover and negatively select against recombinant PrPc. Recombinant PrPc will be expressed from a vector construct containing the elk PrP gene obtained from Dr Katherine ORourke at the USDA via a materials transfer agreement. Characterization of comparative binding of selected aptamers to PrPSc and PrPc will be performed using a radiolabeled aptamer in a dialysis binding assay.
- Medicine and Medical Research
- Environmental Health and Safety