NR ATMA
AU Dabaghian,R.H.; Barnard,G.; Clewley,J.P.
TI A diagnostic assay for the measurement of prion protein in peripheral tissues of CJD patients using differential extraction and DELFIA
QU International Conference - Prion 2005: Between fundamentals and society's needs - 19.10.-21.10.2005, Congress Center Düsseldorf - Poster Session: Diagnosis DIA-64
PT Konferenz-Poster
AB Recent concern about the possible secondary spread of vCJD through blood transfusion and blood products has increased the need for a sensitive and rapid test for the identification of PrPsc in clinical specimens collected in a non-invasive way. In addition, an accurate estimate of the prevalence of pre-clinical vCJD in the British population would be possible if there were such a test that could be applied to readily available material (e.g. blood, urine). As a first step towards this goal, we are developing a simple and sensitive diagnostic test for the detection of PrPsc in the peripheral tissues and brain of vCJD patients, based on time-resolved fluorescence immunoassay (DELFIA) technology. In its present form, the assay uses homogenized tissue from which the PrP isoforms are sequentially extracted by applying two different concentrations of guanidine hydrochloride. Each extraction yields a fraction of the PrP isoforms, which have different solubility. Quantification of the two fractions (insoluble and soluble) using time resolved fluorescence as a reporter system allows differentiation between PrPsc infected and non-infected tissues. This is independent of protein concentration. The assay has a detection limit of 50 pg PrP, is robust and could be automated.
AD R.H.Dabaghian, J.P.Clewley, Virus Reference Department, Centre for Infections, Health Protection Agency, 61 Colindale Avenue, London NW9 5HT, UK; G.Barnard, Centre for Veterinary Science, Department of Clinical Veterinary Medicine
SP englisch
PO Deutschland