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United States Department of Agriculture

Agricultural Research Service

Title: Diagnostic Test Kit for Transmissible Spongiform Encephalopathies Using a New Extraction Method for the Abnormal Prion Protein

Author
item Schmerr, Mary Jo

Submitted to: Patent Application
Publication Type: Abstract Only
Publication Acceptance Date: November 25, 1998
Publication Date: N/A

Technical Abstract: A method to extract the abnormal prion protein from tissues including blood from animals infected with a transmissible spongiform encephalopathy(TSE) was developed using organic solvents such as hexafluoroisopropanol and isopropyl alcohol. The tissues were ground and treated with proteinase K which digests the normal prion protein but not the abnormal prion protein. After digestion, an equal volume of the organic solvent was added to the tissue extract and incubated at 56 deg C for 5 minutes. The layer containing organic solvent was removed and dried in a vacuum centrifuge. The pellet was resuspended in water and applied to a solid phase extraction cartridge. The eluted abnormal prion protein was dried in a vacuum centrifuge and resuspended in water and can be used in many typical immunoassays including dot blots on nitrocellulose paper or ELISA type plate assay or capillary immunoaffinity electrophoresis. This extraction and testing method can be used to diagnose live animals for infection with a TSE. Although this test has been developed primarily for the diagnosis of scrapie (TSE in sheep) it has the potential for diagnosis of other TSEs in other animals and in humans. It also has the potential for use in detection of the abnormal prion protein in process material used to produce human pharmaceuticals or other products intended for human use including food supplements. The test will be of worldwide use in testing for TSE in animals.

Last Modified: 9/22/2014
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