title: Validation of a luminescence immunoassay for the detection of PrP(Sc) in brain homogenate.
abstract: A luminescence immunoassay (LIA) was developed for the diagnosis of bovine spongiform encephalopathy (BSE) in brain tissue using two different monoclonal antibodies for capture and detection of the protease-resistant fragment of the pathological prion protein (PrP27-30). PrP27-30 currently represents the most reliable marker for the infectious particle (denominated prion) causing transmissible spongiform encephalopathies (TSEs). Internal and official validation studies of this assay are described using brain homogenates from ascertained BSE positive and negative cows. Using more than 300 positive and 1400 negative bovine or ovine samples, an excellent sensitivity and specificity of 100% were demonstrated. More than 1000-fold dilutions of a BSE positive homogenate still resulted in a clear positive signal. In combination with a simple homogenisation procedure for the preparation of the samples, this assay lends itself for large scale screening of cattle and sheep for TSEs using complete automation of the process.
authors.affiliation: Prionics AG, University of Zurich, Winterthurerstr. 190, CH-8057, Zurich, Switzerland.
authors.forename: Karin
authors.initials: K
authors.lastname: Biffiger
authors.forename: Daniel
authors.initials: D
authors.lastname: Zwald
authors.forename: Lukas
authors.initials: L
authors.lastname: Kaufmann
authors.forename: Alexandra
authors.initials: A
authors.lastname: Briner
authors.forename: Inci
authors.initials: I
authors.lastname: Nayki
authors.forename: Mario
authors.initials: M
authors.lastname: Pürro
authors.forename: Sigrid
authors.initials: S
authors.lastname: Bottcher
authors.forename: Thomas
authors.initials: T
authors.lastname: Struckmeyer
authors.forename: Olivier
authors.initials: O
authors.lastname: Schaller
authors.forename: Rudolf
authors.initials: R
authors.lastname: Meyer
authors.forename: Rosemarie
authors.initials: R
authors.lastname: Fatzer
authors.forename: Andres
authors.initials: A
authors.lastname: Zurbriggen
authors.forename: Mick
authors.initials: M
authors.lastname: Stack
authors.forename: Markus
authors.initials: M
authors.lastname: Moser
authors.forename: Bruno
authors.initials: B
authors.lastname: Oesch
authors.forename: Eric
authors.initials: E
authors.lastname: Kübler
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015415:Biomarkers / Q000032:analysis; D001923:Brain Chemistry; D002417:Cattle; D002457:Cell Extracts / Q000032:analysis; D016643:Encephalopathy, Bovine Spongiform / Q000175:diagnosis* / Q000453:epidemiology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D008163:Luminescent Measurements; D051379:Mice; D011159:Population Surveillance; D018622:PrP 27-30 Protein / Q000032:analysis* / Q000302:isolation & purification; D015203:Reproducibility of Results; D012608:Scrapie / Q000175:diagnosis; D012680:Sensitivity and Specificity; D012756:Sheep; D013557:Switzerland / Q000453:epidemiology
chemical_list: D000911:Antibodies, Monoclonal; D015415:Biomarkers; D002457:Cell Extracts; D018622:PrP 27-30 Protein