NR ATLE
AU Simon,S.; Nugier,J.; Boutal,H.; Benestad,S.L.; Andreoletti,O.; Feyssaguet,M.; Bilheude,J.M.; Biacabe,A.G.; Baron,T.G.M.; Grassi,J.
TI Rapid Typing of TSE strains using a differential ELISA technique. Application to the identification of BSE in small ruminant flocks.
QU International Conference - Prion 2005: Between fundamentals and society's needs - 19.10.-21.10.2005, Congress Center Düsseldorf - Poster Session: Diagnosis DIA-42
PT Konferenz-Poster
AB
Several lines of evidence have shown that the bovine spongiform encephalopathy (BSE) agent has been transmitted to human via the food chain, leading to the variant Creutzfeldt-Jakob disease (vCJD). As sheep and goats can be experimentally infected by BSE and have been potentially exposed to BSE through the consumption of meat and bone meal, the question of the transmission of BSE to small ruminants has been raised for many years.
Screening for BSE in sheep and goats is therefore a priority in research into transmissible spongiform encephalopathies (TSE). Based on the particular properties of the abnormal prion protein (PrPres) and the possible discrimination of BSE and scrapie strains by proteinase K treatment, we have developed an high throughput ELISA based technique designed to distinguish BSE from other scrapie strains. Each sample is processed using two sets of conditions for PK treatment and absorbance ratios obtained by the ELISA in the two conditions are measured and normalized with the ratio obtained for the internal ovine BSE control. This test can very clearly discriminate experimental BSE in sheep from other scrapie strains and has been used to screen more than four hundred and fifty TSE infected ovine samples identified by the French active surveillance network between 1996 and 2003. On this occasion it has participated to the identification of the first case of natural BSE in a goat. In addition, we show that this approach allows classifying TSE isolates according to the PK susceptibility of the associated PrPres. As a consequence, it also provides a direct identification of atypical scrapie cases characterized by a PrPres bearing an unusually high susceptibility to PK degradation, including animals infected by the Nor-98 strain. The test is also suitable to analyse the PK resistance of CWD and CJD infected brain samples.
IN Die Autoren untersuchten mehr als 450 Proben von Scrapie-infizierten Schafen, die zwischen 1996 und 2003 im Rahmen des französischen Scrapie-Überwachungsprogramms identifiziert wurden. Dabei teilten sie sämtliche Proben und führten mit den beiden Hälften jeweils zwei unterschiedlich intensive Behandlungen mit der Proteinase K durch. Die beiden unterschiedlichen Protokolle zur Proteinase-K-Verdauung hatten unterschiedliche ELISA-Signalstärken zur Folge, deren Quotienten offenbar eine gute Unterscheidung zwischen konventionellen, atypischen und durch BSE verursachte Scrapie-Fällen erlauben. Mit dieser Methode fanden die Autoren auch den ersten bekannten Fall einer BSE-infizierten Ziege in der normalen Landwirtschaft.
AD Stéphanie Simon, Jérôme Nugier, Hervé Boutal, Jacques Grassi, CEA, Service de Pharmacologie et d'Immunologie, CEA/Saclay, 91191 Gif sur Yvette, France; Sylvie Benestad, National Veterinary Institute, PO Box 8156 Dep., N-0033 Oslo, Norway; Olivier Andréoletti, UMR INRA ENVT 1225, Interactions Hôte Agent Pathogène, Ecole Nationale Vétérinaire de Toulouse, 23 Chemin des Capelles, 31076 Toulouse, France.; Muriel Feyssaguet, Jean Marc Bilheude, Bio-rad, 92430 Marnes La Coquette, France; Anne Gaëlle Biacabe, Thierry Baron, AFSSA-Lyon, Unité 'Virologie-ATNC', 31 Avenue Tony Garnier, 69364 Lyon, France.
SP englisch
PO Deutschland