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M03034: Detection of BSE prions from cattle and sheep using transgenic mice and conformation-dependent immunoassay

Wednesday 10 September 2003

This research project aims to improve methods for the detection of BSE infection in cattle and sheep including differentiation between BSE and scrapie infection in sheep.

Study Duration : September 2003 to August 2006

Contractor : University of California, San Francisco

Background

The aim of this joint FSA/ Defra study is to further characterise two complementary methods for detection of and discrimination between BSE and scrapie prions. These two methods are bioassay using transgenic mice expressing bovine prion protein (Tg(BoPrP) and the in vitro Confirmation-Dependent Immunoassay (CDI).

Research Approach

Calibration studies will be carried out for the detection of BSE and scrapie prions using bioassay with Tg(BoPrP) mice. The sensitivity of these Tg mice to BSE and scrapie prions will be established by endpoint titration of standardized BSE stock inocula and the data will be used to develop an incubation-time bioassay. The same inocula are to be tested in parallel in RIII mice (an established bioassay system), to effect a direct comparison of the relative sensitivities of transgenic mice and inbred mice to BSE and scrapie prions.

In addition, the CDI will be evaluated and calibrated. The immunoassay data will be correlated with bioassay data and statistically evaluated for diagnostic sensitivity, specificity, and both positive and negative predictive value. Finally, it is proposed that both the CDI and bioassay in Tg mice will be validated for the detection of BSE prions in sheep and the discrimination between BSE and scrapie prions to address the issue of whether BSE prions may have been passed to sheep in the UK.

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