Optimizing Fecal Sample Preparation to Determine Presence of Chronic Wasting Disease in White Tail Deer
presentationposted on 21.07.2014, 00:00 by Aya Murakami
Chronic Wasting Disease (CWD) is a prion disease that affects cervids (deer, elk, and moose). A prion disease is characterized by the accumulation of a misfolded isoform of the prion protein and is invariably fatal to the host. Currently, diagnosis of CWD can be done only with post mortem testing of animal tissues or recto-anal mucosa associated lymphoid tissue (RAMALT) analysis, by the direct detection of prion protein. The purpose of this project is to develop a fecal test for CWD biomarkers to enable us to have a simple ante mortem test. Gentle feces surface extraction was performed to obtain the proteins present in the mucous layer surrounding a deer pellet. Western blots with decreased background were observed when extraction was done in fresh buffer with appropriate amount of protease inhibitors. Background noise decreased as extraction time was shortened and the shaking became gentler; whereas, the concentration of primary and secondary antibodies had less effect on over all background. Good progress has been made toward perfecting fecal extraction to detect biomarkers for CWD.