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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: Diagnosing Prion Diseases: Mass Spectrometry-Based Approaches

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

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: May 11, 2011
Publication Date: May 16, 2011
Citation: Silva, C.J., Erickson, M.L., Dynin, I.A., Onisko, B.C., Carter, J.M. 2011. Diagnosing Prion Diseases: Mass Spectrometry-Based Approaches. Meeting Abstract. Prion, Volume 5 (Supplement), April/May/June 2011. page 85.

Technical Abstract: Mass spectrometry is an established means of quantitating the prions present in infected hamsters. Calibration curves relating the area ratios of the selected analyte peptides and their oxidized analogs to stable isotope labeled internal standards were prepared. The limit of detection (LOD) and limit of quantitation (LOQ) for human, sheep, deer, cow, and mouse PrP were determined to be below 100 attomoles. Non-analyte peptides that were characteristic of prions were included in the multiple reaction monitoring method. These peptides had LODs much lower than those of the analyte peptides, thereby allowing for both the quantitation and confirmation of the presence of prions in the attomole range. This method was used to quantitate the prions present in brains of hamsters or mice five weeks after inoculation (ic) with either four hamster-adapted prion strains or four mouse-adapted prion strains. The prions from different brain regions of a sheep naturally infected with scrapie were also quantitated. All of the rodent-adapted prion strains were detectable in the asymptomatic animals. In sheep, prions were detectable in the obex, anterior portion of the cerebrum, and the non-obex/non-anterior portion of the cerebrum. This mass spectrometry-based approach can be used to quantitate and confirm the presence of prions before detectable pathology.

Last Modified: 10/1/2014
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