The measurement of prion protein in bovine brain tissue using differential extraction and DELFIA� as a diagnostic test for BSE
2000; Wiley; Volume: 15; Issue: 6 Linguagem: Inglês
10.1002/1522-7243(200011/12)15
ISSN1522-7243
AutoresGeoff Barnard, Brett Helmick, Sean Madden, Chris Gilbourne, Raj Patel,
Tópico(s)Neurological diseases and metabolism
ResumoAn 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.
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