In vitro amplification of H-type atypical bovine spongiform encephalopathy by protein misfolding cyclic amplification

O'Connor, M.J., Bishop, Keith, Workman, Robert G., Maddison, Ben C. and Gough, Kevin C. (2017) In vitro amplification of H-type atypical bovine spongiform encephalopathy by protein misfolding cyclic amplification. Prion, 11 (1). pp. 54-64. ISSN 1933-690X

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Abstract

The in vitro amplification of prions by serial protein misfolding cyclic amplification has been shown to detect PrPSc to 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 three distinct forms, classical BSE (C-BSE) and two atypical BSE forms (L-BSE and H-BSE). Atypical forms are usually detectedin 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 PrPSc from 3 distinct experimental isolates of H-BSE, could detect PrPSc in as little as 1 x 10-12 g of brain material and was highly specific. Additionally, the product of serial protein misfolding cyclic amplification at all dilutions of seed analysed could be readily distinguished from L-BSE, which did not amplify, and C-BSE, which had PrPSc with distinct protease K-resistance and protease K-resistant PrPSc molecular weights.

Item Type: Article
RIS ID: https://nottingham-repository.worktribe.com/output/846843
Keywords: Atypical prions, Bovine spongiform encephalopathy, H-type BSE, PMCA
Schools/Departments: University of Nottingham, UK > Faculty of Medicine and Health Sciences > School of Veterinary Medicine and Science
Identification Number: https://doi.org/10.1080/19336896.2016.1259051
Depositing User: Eprints, Support
Date Deposited: 13 Jan 2017 08:45
Last Modified: 04 May 2020 18:35
URI: https://eprints.nottingham.ac.uk/id/eprint/39831

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