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Creation of a bench test that is sensitive, specific, reproducible and ante-mortem for detection of TSE infection specifically CWD, be it in the seemingly healthy individual or animal. Current tests are either ante-mortem or use tissues that are difficult to obtain. This research is designed to eliminate both problems by looking at surrogate protein markers not the prion protein itself.
NON-TECHNICAL SUMMARY: Transmissible Spongiform Encephalopathy (TSE) diseases such as BSE and CWD have proved notoriously difficult to detect in the live animal. Current diagnostic methods, and some under development, require brain tissue from the animal to be tested and require days to be completed. The U.S. slaughters 37 million head of cattle per year making testing quite difficult when the results cannot be obtained for days. Past and current attempts to detect the prion, the infectious agent of TSE disease, have been met with considerable obstacles and are not likely a near term possibility. Considering the obstacles of detecting the prion, our group has pursued and identified nine different protein biomarkers using chronic wasting disease for test development, that can be detected to accurately diagnose the disease. These proteins, much like other known biomarkers such as human chorionic gonadotropin and prostrate specific antigen, are present when the disease is present and are not detectableat other times. Protein biomarkers show tremendous potential for detecting TSE's in that the test formats can be made simple enough to be performed by the lay person in addition to being animal side rather than laboratory based providing an answer within minutes to the slaughter house attendant or the sportsman in the field. This project is designed to validate these markers and test them on urine from infected and control animals.
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APPROACH: Work to accomplish the technical objective will progress with these tasks:<OL> <LI> PCR and sequence the deer specific-forms of the nine proteins. <LI> Identify peptide sequences in common and synthesizepeptides. <LI> Generate polyclonal antibodies to the selected peptide sequences. <LI> Validate antibodies, and thus biomarkers, using infected and non-infected urine samples to establish their feasibility as protein biomarkers of the disease. <LI> Accomplishment of 5. will lead to the next step of development of the assay and field testing which will be beyond the time scope of this project.