NR AAZP
AU Barnard,G.; Helmick,B.; Madden,S.; Gilbourne,C.; Patel,R.
TI The measurement of prion protein in bovine brain tissue using differential extraction and DELFIA as a diagnostic test for BSE
QU Luminescence : The Journal of Biological and Chemical Luminescence 2000 Nov-Dec; 15(6): 357-62
ER Luminescence 2001 Mar-Apr;16(2):211
PT journal article
AB A simple diagnostic test for the detection of bovine spongiform encephalopathy (BSE), based on a commercially available time-resolved fluorescence immunoassay (DELFIA) for the measurement of the normal and disease-associated isoforms of prion protein (PrP), is described. The isoforms are sequentially extracted from homogenized bovine brain tissue using two concentrations of guanidine hydrochloride. This procedure initially extracts a soluble isoform and subsequently a less soluble disease-associated aggregated isoform. Following quantification of the two fractions, the percentage of the insoluble prion becomes a measurable parameter, independent of protein concentration, clearly identifying normal from infected animals displaying clinical signs of BSE. The mean percentages of insoluble PrP in brain tissue from 60 BSE-confirmed-positive cattle and 100 cattle that had never been exposed to the disease were 52.6% (SD = 22.8) and 3.9% (SD = 1.5), respectively. The assay is sensitive, with a detection limit of less than 50 pg PrP, and is robust and precise (CVs < 10%) over the appropriate working range.
MH Animal; *Brain Chemistry; Cattle; Encephalopathy, Bovine Spongiform/*diagnosis; Endopeptidase K; Fluoroimmunoassay/methods/standards/*veterinary; Prions/*analysis/isolation & purification; Solubility; Support, Non-U.S. Gov't
AD Endocrine Unit, Department of Chemical Pathology, Level D, South Pathology Block, Southampton General Hospital, Tremona Road, Southampton SO16 6YD, UK. GJRB2@aol.com
SP englisch
PO England