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Research Paper
Direct Detection of Disease-Associated Prions in Brain and Lymphoid Tissue Using Antibodies Recognizing the Extreme N-terminus of PrPC
Geoff Barnard, Lee Hopkins, Sowmiya Moorthie, David Seilly, Paul Tonks, Reza Dabaghian, Jonathan Clewley, John Coward and Ian McConnell
volume 1 | issue 2
April/May/June 2007Pages: 121 - 127
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A simple diagnostic test is described for the detection of TSE in bovine, ovine and human brain and lymphoid tissue that obviates the use of proteinase K as a discriminating reagent. The immunoassay utilises high affinity anti-peptide antibodies that appear blind to the normal isoform of prion protein (PrPC). These reagents have been produced with novel N-terminal chimeric peptides and we hypothesise that the retention and stability of the extreme N-terminus of PrP in the disease-associated aggregate makes it an operationally specific marker for TSE. Accordingly, the assay involves homogenisation of the tissue directly in 8M guanidine hydrochloride, a simple one-step capture of PrPSc followed by detection with a europium-labelled anti-PrPC antibody. This rapid assay clearly differentiates between levels of disease-associated PrP extracted from brain and lymphoid tissues taken from confirmed TSE positive and negative cattle and sheep. The assay can also be used to detect PrPSc in cases of vCJD.
Authors
Geoff Barnard
Centre for Veterinary Science, Department of Veterinary Medicine, University of Cambridge, Cambridge UK
Lee Hopkins
Centre for Veterinary Science, Department of Veterinary Medicine, University of Cambridge, Cambridge UK
Sowmiya Moorthie
Centre for Veterinary Science, Department of Veterinary Medicine, University of Cambridge, Cambridge UK
David Seilly
Centre for Veterinary Science, Department of Veterinary Medicine, University of Cambridge, Cambridge UK
Paul Tonks
Centre for Veterinary Science, Department of Veterinary Medicine, University of Cambridge, Cambridge UK
Reza Dabaghian
Virus Reference Department, Health Protection Agency, London UK
Jonathan Clewley
Virus Reference Department, Health Protection Agency, London UK
John Coward
RevelationBio Ltd., Dorset UK
Ian McConnell
Centre for Veterinary Science, Department of Veterinary Medicine, University of Cambridge, Cambridge UK
We now provide open access to journal articles published online for one year or more. This article may be downloaded at the following link:
Download PDF If the document does not open, please right-click on the link (control-click on a Macintosh) and select the option to save the file to disk.