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

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 i...

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Main Authors: O'Connor, M.J., Bishop, Keith, Workman, Robert G., Maddison, Ben C., Gough, Kevin C.
Format: Article
Published: Taylor & Francis 2017
Subjects:
Online Access:https://eprints.nottingham.ac.uk/39831/
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author O'Connor, M.J.
Bishop, Keith
Workman, Robert G.
Maddison, Ben C.
Gough, Kevin C.
author_facet O'Connor, M.J.
Bishop, Keith
Workman, Robert G.
Maddison, Ben C.
Gough, Kevin C.
author_sort O'Connor, M.J.
building Nottingham Research Data Repository
collection Online Access
description 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.
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spelling nottingham-398312020-05-04T18:35:36Z https://eprints.nottingham.ac.uk/39831/ 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. 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. Taylor & Francis 2017-02-08 Article PeerReviewed 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 Atypical prions Bovine spongiform encephalopathy H-type BSE PMCA http://www.tandfonline.com/doi/full/10.1080/19336896.2016.1259051 doi:10.1080/19336896.2016.1259051 doi:10.1080/19336896.2016.1259051
spellingShingle Atypical prions
Bovine spongiform encephalopathy
H-type BSE
PMCA
O'Connor, M.J.
Bishop, Keith
Workman, Robert G.
Maddison, Ben C.
Gough, Kevin C.
In vitro amplification of H-type atypical bovine spongiform encephalopathy by protein misfolding cyclic amplification
title In vitro amplification of H-type atypical bovine spongiform encephalopathy by protein misfolding cyclic amplification
title_full In vitro amplification of H-type atypical bovine spongiform encephalopathy by protein misfolding cyclic amplification
title_fullStr In vitro amplification of H-type atypical bovine spongiform encephalopathy by protein misfolding cyclic amplification
title_full_unstemmed In vitro amplification of H-type atypical bovine spongiform encephalopathy by protein misfolding cyclic amplification
title_short In vitro amplification of H-type atypical bovine spongiform encephalopathy by protein misfolding cyclic amplification
title_sort in vitro amplification of h-type atypical bovine spongiform encephalopathy by protein misfolding cyclic amplification
topic Atypical prions
Bovine spongiform encephalopathy
H-type BSE
PMCA
url https://eprints.nottingham.ac.uk/39831/
https://eprints.nottingham.ac.uk/39831/
https://eprints.nottingham.ac.uk/39831/