Direct detection of disease associated prions in brain and lymphoid tissue using antibodies recognizing the extreme N terminus of PrPC.
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 (PrP(C)). 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 PrP(Sc) followed by detection with a europium-labelled anti-PrP(C) 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 PrP(Sc) in cases of vCJD.
Barnard
Geoff
G
Department of Veterinary Medicine, University of Cambridge, Cambridge, UK. gjrb3@cam.ac.uk
Hopkins
Lee
L
Moorthie
Sowmiya
S
Seilly
David
D
Tonks
Paul
P
Dabaghian
Reza
R
Clewley
Jonathan
J
Coward
John
J
McConnell
Ian
I
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D000918:Antibody Specificity; D001921:Brain / Q000276:immunology; D001923:Brain Chemistry; D002417:Cattle; D006801:Humans; D008221:Lymphoid Tissue / Q000737:chemistry* / Q000276:immunology; D018621:PrPC Proteins / Q000737:chemistry* / Q000276:immunology; D018620:PrPSc Proteins / Q000737:chemistry* / Q000276:immunology; D017096:Prion Diseases; D012756:Sheep
D000911:Antibodies, Monoclonal; D018621:PrPC Proteins; D018620:PrPSc Proteins