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.; Gough, Kevin C.

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.

Citation

O'Connor, M., Bishop, K., Workman, R. G., Maddison, B. C., & Gough, K. C. (in press). In vitro amplification of H-type atypical bovine spongiform encephalopathy by protein misfolding cyclic amplification. Prion, 11(1), 54-64. https://doi.org/10.1080/19336896.2016.1259051

• Journal Article Type: Article
• Acceptance Date: Nov 4, 2016
• Online Publication Date: Feb 8, 2017
• Deposit Date: Jan 13, 2017
• Publicly Available Date: Feb 8, 2017
• Journal: Prion
• Print ISSN: 1933-6896
• Electronic ISSN: 1933-690X
• Publisher: Taylor and Francis
• Peer Reviewed: Peer Reviewed
• Volume: 11
• Issue: 1
• Pages: 54-64
• DOI: https://doi.org/10.1080/19336896.2016.1259051
• Keywords: Atypical prions, Bovine spongiform encephalopathy, H-type BSE, PMCA
• Public URL: https://nottingham-repository.worktribe.com/output/846843
• Publisher URL: http://www.tandfonline.com/doi/full/10.1080/19336896.2016.1259051
• Additional Information: Peer Review Statement: The publishing and review policy for this title is described in its Aims & Scope.; Aim & Scope: http://www.tandfonline.com/action/journalInformation?show=aimsScope&journalCode=kprn20
• Contract Date: Jan 13, 2017