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United States Department of Agriculture

Agricultural Research Service

Research Project: NOVEL STRATEGIES FOR THE DETECTION OF INFECTIOUS PRION PROTEINS
2009 Annual Report


1a.Objectives (from AD-416)
To develop new reagents and methods for the detection and evaluation of prion proteins in animal and environmental samples.


1b.Approach (from AD-416)
Bovine Spongiform Encephalopathy (BSE) and other transmissible prion diseases represent an important agricultural issue and pose unique diagnostic challenges. Unlike conventional microbes, prions do not require agent-specific nucleic acid in order to multiply. Propagation occurs in infected animals when normal prion protein (PrPc) becomes “misfolded” into an infectious form (PrPsc) in a template-driven process. No PrPsc specific probes have been developed and conventional methods rely on differences in the sensitivity of PrPc from PrPsc to chemical reagents for detection. BSE has had a devastating impact on foreign agricultural economies and despite a U.S. agricultural ban on ruminant protein imports from these countries, BSE remains a bio-security threat. New methods for the sensitive detection of PrPsc in animals, their by-products and the environment are essential for preventing the transmission of disease. We will develop strategies for the extraction and enrichment of PrPsc for use in the development of novel diagnostic reagents and platforms. These models will be used for the assessment of low-level PrPsc in pre-clinical animals and the environment. Methods will focus on isolation of PrPsc in detergent resistant membranes, utilization of sensitive transgenic mice as models of infectivity, cell based bioassay, and production of monoclonal antibodies. The generation and establishment of new reagents and detection platforms will provide the necessary diagnostic tools to achieve early detection of low-level PrPsc in contaminated biological and environmental samples. Documents SCA with UCSF. Formerly 5325-32000-007-02S (4/08).


3.Progress Report

Previously in this project we developed a strain of “knockout” mouse, which does not have the PrP protein known to be necessary for development of prion diseases. This year we inoculated some of these mice (as well as normal “wild-type” mice) with hamster prion from our new sample preparation method. Although the wild type animals did not mount an immune response, our knockout mice gave a robust antibody reaction. We used these animals to generate monoclonal antibodies and isolated five new high-affinity anti-prion monoclonal antibodies.

ADODR monitors this project through occasional visits, telephone conversations, and email with the Cooperator. ADODR and Cooperator conduct annual group meetings with all personnel, and occasionally meet at national scientific meetings for additional coordination.


Last Modified: 10/24/2014
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