NR ATGR
AU Perron,H.; Dupin,M.; Cecillon,S.; Jolivet,M.; Kopp,N.; Chazot,G.; Vighetto,A.; Moussa,A.; Coleman,A.W.; Perret-Liaudet,A.
TI A novel immunoassay for the detection of resistant prion protein (PrPres) in human plasma
QU International Conference - Prion 2005: Between fundamentals and society's needs - 19.10.-21.10.2005, Congress Center Düsseldorf - Oral sessions ORAL-48
PT Konferenz-Vortrag
AB
Transmission of infectious Prion protein by blood product transfusion from asymptomatic carriers to human recipients was evidenced by recent reports of UK patients who developed nvCJD or were infected by nvCJD agent after blood transfusion from asymptomatic donors who developed nvCJD after blood donation. Epidemiological studies also indicated that few thousands of asymptomatic carriers of nvCJD agent exist within UK population. Thus the detection of PrPres in the blood products of asymptomatic carriers before they ever develop clinical signs of CJD, has become a critical unmet need for blood transfusion safety.
Targeting the detection of pathogenic PrP by its specific proteinase-K (PK) resistant fragment (PrPres) in human and animal blood, we have evaluated molecules capable of binding to PrP. Streptomycine, under certain physico-chemical conditions, was found to aggregate PrPres in BSE or CJD brain samples after PK digestion, as well as in purified Scrapie Associated Fibrils from different species. Another molecule, Calix-6-Arene sulfonate, was found to bind efficiently to PrPres aggregated by streptomycine, due to its "molecular basket" structure.
A microplate immunoassay prototype for the detection of PrPres in human blood has been designed using the combined properties of these two molecules, with final specific antigenic detection using an anti-PrP monoclonal (8D11G12, bioMérieux).
"Heparin" plasma from 10 CJD, including sporadic type 1 and 2, new variants and Iatrogenic cases, and 500 french blood donors were tested. 9/10 CJD plasma were positive but none of the 500 healthy blood donors. Beyond this proof of principle of PrPres detection in human blood, detection in asymptomatic carriers and a test for excluding blood donations from such apparently healthy donors are next objectives. Further studies and technical improvements are required before it may be proposed for validation as a routine test for blood banking and/or human CJD diagnosis.
IN Auf der Grundlage der Bindung von Streptomycin an PrPres und der Bindung von Calix-6-Aren-Sulfonat an Komplexe von PrPres und Streptomycin entwickelten die Autoren einen CJK-Bluttest, mit welchem sie 9 von 10 CJK-Patienten anhand ihrer Plasmaproben identifizierten, während es bei 500 gesunden Kontrollpersonen zu keinem einzigen falsch-positiven Ergebnis kam.
AD H.Perron, M.Dupin, M.Jolivet, bioMerieux, R&D Immunoassay and proteomics, Lyon-France.; S.Cecillon, A.W.Coleman, Institut de Biologie te de Chimie des proteines, CNRS, Lyon-France; A.Moussa, Agence Française pour la Sécurité Sanitaire des Aliments, Lyon-France; N.Kopp, G.Chazot, A.Vighetto4, A.Perret-Liaudet, Federation des maladies a Prions, Hopital neurologique, Lyon-France
SP englisch
PO Deutschland