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 (Prp(Sc)). 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 PrP(Sc) but not in PrP(C). Next, selection will target CWD PrP(Sc) using a novel electrodialysis SELEX technique being developed in our laboratory. The final aptamer selection will crossover and negatively select against recombinant PrP(c). Recombinant PrP(c) will be expressed from a vector construct containing the elk PrP gene obtained from Dr Katherine O'Rourke at the USDA via a materials transfer agreement. Characterization of comparative binding of selected aptamers to PrP(Sc) and PrP(c) will be performed using a radiolabeled aptamer in a dialysis binding assay.
Document Details
- Document Type
- Technical Report
- Publication Date
- May 01, 2005
- Accession Number
- ADA446338
Entities
People
- Kenneth D. Clinkenbeard
Organizations
- Oklahoma State University–Stillwater