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United States Department of Agriculture

Agricultural Research Service

Research Project: DETECTION OF TRANSMISSIBLE SPONGIFORM ENCEPHALOPATHY AGENTS IN LIVESTOCK, WILDLIFE, AGRICULTURAL PRODUCTS, AND THE ENVIRONMENT Title: Mass spectrometric approaches to detecting prions and protein conformers

Authors
item Silva, Christopher
item Onisko, Bruce
item Dynin, Irina
item Erickson-Beltran, Melissa

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: September 21, 2008
Publication Date: September 22, 2009
Citation: Silva, C.J., Onisko, B.C., Dynin, I.A., Erickson, M.L. 2009. Mass spectrometric approaches to detecting prions and protein conformers. [Abstract] AOAC S-401. p63

Technical Abstract: Transmissible spongiform encephalopathies (TSEs) can cause substantial economic damage to agriculture. These diseases have characteristically long incubation periods, comparatively short symptomatic intervals, and are invariably fatal. Early detection is important in controlling these diseases. However, detecting prions is a challenging endeavor, for a single infectious dose is estimated to be 100,000 to 1,000,000 molecules or 1 attomole (10(exp-18) mole). Furthermore, the only differences between the infectious prion and the normal cellular prion protein (PrPC) are conformational. A given host can propagate more than one kind of prion, referred to as a strain. These strains have distinct properties, which need to be distinguished. We have developed a mass spectrometry (MS) based approach to detect prions. This approach can be used to detect prions in experimental rodents, sheep, deer, elk, cows and humans. Our approach uses detergent solubility and proteinase K to eliminate the PrPC. The prion is inactivated before MS analysis, so the instrument does not become contaminated. This method is superior to any other instrumental technique and unlike other methods this approach provides for a direct detection of the prion protein. This method is comparable to the most sensitive bioassay.

Last Modified: 12/19/2014
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