Aptamer selection and cell culture model development for diagnosis of chronic wasting disease of Cervidae

Chronic wasting disease (CWD) is a transmissible spongiform encephalopathy that affects cervid species (deer, elk, and moose) in both captive and wild animals. Since first identified and described in the late 1970's, CWD has had significant impact upon the captive cervid industry and has been found in an increasingly large geographic area of North America in wild populations. The association of bovine spongiform encephalopathy (a similar disease that occurs in cattle) with a new form of human encephalopathy in 1986 caused a dramatic increase in awareness and concern about all transmissible encephalopathies, including CWD. This led to an increase in surveillance efforts for CWD that may account for part of the expanding incidence that has been reported in the last twenty years. Although there are no reports of zoonotic disease associated with CWD and there are lines of evidence that show transmission to humans is unlikely, the infectious agent has recently been identified in skeletal muscle of mule deer and the exact nature of the infectious agent is still poorly understood.; One of the most complex problems associated with CWD and other transmissible spongiform encephalopathies is the lack of diagnostic methods to identify preclinical cases of disease. The studies described herein are focused on solving this difficult problem using some novel approaches as compared to development of diagnostic strategies for other infectious agents. The first study is a discovery based experiment designed to select a nucleic acid recognition ligand, and the second is designed to develop a model of CWD in cell culture that could serve as a very sensitive detection system.; Both of these studies were high risk endeavors attempting to solve the diagnostic dilemma of prion diseases with some very novel approaches. The identification of one aptamer that shows some specificity for PrPCWD only opens the door for further characterization and adaptation of this potential ligand. The results of the cell culture model development raise some stimulation questions about prion protein trafficking and control, and served as a basis for the development of other cell culture models.