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The overall objective of Phase II is to develop an in vitro culture system to amplify Chronic Wasting Disease (CWD) prions from samples of blood, brain, lymph node, and other samples from affected animals. Once defined, the utility of the system to differentiate between prion species and strains will be established.
APPROACH:<ol>
<li> Confirm that deer and elk Follicular Dendritic Cells (FDCs) are permissive to PrP**res replication similar to the ovine system. <li> Optimize the assay for maximum efficiency by determining minimal parameters for PrP**Sc replication.<li> Define the sensitivity of the in vitro bioassay by direct comparison with established diagnostic assays including Western Blot and Immunohistochemistry.<li> Define the utility of the assay for diagnosis of specific tissue samples from Chronic Wasting Disease (CWD)-infected deer and elk. <li> Investigate the use of FDC-B cell cultures to define the species barrier of CWD. </ol>