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The measurement of prion protein in bovine brain tissue using differential extraction and DELFIA® as a diagnostic test for BSE
Author(s) -
Barnard Geoff,
Helmick Brett,
Madden Sean,
Gilbourne Chris,
Patel Raj
Publication year - 2000
Publication title -
luminescence
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.428
H-Index - 45
eISSN - 1522-7243
pISSN - 1522-7235
DOI - 10.1002/1522-7243(200011/12)15:6<357::aid-bio621>3.0.co;2-v
Subject(s) - bovine spongiform encephalopathy , gene isoform , prion protein , detection limit , guanidine , microbiology and biotechnology , brain tissue , immunoassay , chemistry , pathology , biology , virology , disease , medicine , chromatography , biochemistry , immunology , antibody , gene
An Erratum has been published for this article in Luminescence 2001;(16):211 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. Copyright © 2000 John Wiley & Sons, Ltd.