title: In vitro amplification of H-type atypical bovine spongiform encephalopathy by protein misfolding cyclic amplification.
abstract: The in vitro amplification of prions by serial protein misfolding cyclic amplification has been shown to detect PrPScto levels at least as sensitive as rodent bioassay but in a fraction of the time. Bovine spongiform encephalopathy is a zoonotic prion disease in cattle and has been shown to occur in 3 distinct forms, classical BSE (C-BSE) and 2 atypical BSE forms (L-BSE and H-BSE). Atypical forms are usually detected in asymptomatic, older cattle and are suggested to be spontaneous forms of the disease. Here, we show the development of a serial protein misfolding cyclic amplification method for the detection of H-BSE. The assay could detect PrPScfrom 3 distinct experimental isolates of H-BSE, could detect PrPScin as little as 1×10-12g of brain material and was highly specific. Additionally, the product of serial protein misfolding cyclic amplification at all dilutions of seed analyzed could be readily distinguished from L-BSE, which did not amplify, and C-BSE, which had PrPScwith distinct protease K-resistance and protease K-resistant PrPScmolecular weights.
authors.affiliation: a School of Veterinary Medicine and Science, The University of Nottingham, Sutton Bonington Campus , College Road, Sutton Bonington , Leicestershire , UK.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: O'Connor
authors.affiliation: b ADAS UK, School of Veterinary Medicine and Science, The University of Nottingham, Sutton Bonington Campus , College Road, Sutton Bonington , Leicestershire , UK.
authors.forename: Keith
authors.initials: K
authors.lastname: Bishop
authors.affiliation: a School of Veterinary Medicine and Science, The University of Nottingham, Sutton Bonington Campus , College Road, Sutton Bonington , Leicestershire , UK.
authors.forename: Robert G
authors.initials: RG
authors.lastname: Workman
authors.affiliation: b ADAS UK, School of Veterinary Medicine and Science, The University of Nottingham, Sutton Bonington Campus , College Road, Sutton Bonington , Leicestershire , UK.
authors.forename: Ben C
authors.initials: BC
authors.lastname: Maddison
authors.affiliation: a School of Veterinary Medicine and Science, The University of Nottingham, Sutton Bonington Campus , College Road, Sutton Bonington , Leicestershire , UK.
authors.forename: Kevin C
authors.initials: KC
authors.lastname: Gough
mesh_terms: D000818:Animals; D002417:Cattle; D016643:Encephalopathy, Bovine Spongiform / Q000378:metabolism*; D000072002:Prion Proteins / Q000378:metabolism*
chemical_list: D000072002:Prion Proteins
keywords: H-type BSE; PMCA; atypical prions; bovine spongiform encephalopathy