A sensitive 301V BSE serial PMCA assay

The prion strain 301V, is a mouse passaged form of bovine spongiform encephalopathy (BSE). It has been used as a model of BSE for more than 20 years, in particular in the investigation of tissue distribution of infectivity, the molecular phenotype and transmission properties of BSE, strain typing as...

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Main Authors: Gough, Kevin C., Bishop, Keith, Somerville, Robert A, Hunter, Nora, Maddison, Ben C.
Format: Article
Published: F1000Research 2016
Online Access:https://eprints.nottingham.ac.uk/38513/
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author Gough, Kevin C.
Bishop, Keith
Somerville, Robert A
Hunter, Nora
Maddison, Ben C.
author_facet Gough, Kevin C.
Bishop, Keith
Somerville, Robert A
Hunter, Nora
Maddison, Ben C.
author_sort Gough, Kevin C.
building Nottingham Research Data Repository
collection Online Access
description The prion strain 301V, is a mouse passaged form of bovine spongiform encephalopathy (BSE). It has been used as a model of BSE for more than 20 years, in particular in the investigation of tissue distribution of infectivity, the molecular phenotype and transmission properties of BSE, strain typing assays and prion inactivation studies. Most 301V experiments have required murine bioassay as a method for the quantitation of infectivity. To date this model strain has not been studied with the protein misfolding cyclic amplification assay (PMCA) which detects prion-associated PrPSc protein. The detection of BSE PrPSc by PMCA can be more sensitive than mouse bioassay and is carried out in a much shorter time frame of days as opposed to months/years. Here, we describe the development of a new highly sensitive and specific PMCA assay for murine 301V and assess the sensitivity of the assay in direct comparison with murine bioassay of the same material. This in vitro assay detected, in a few days, 301V at a brain dilution of at least 1x10-9, compared to bioassay of the same material in VM mice that could detect down to a 1x10-8 dilution and took >180 days. The 301V PMCA may therefore offer a faster and more sensitive alternative to live animal bioassay when studying the BSE agent in VM mice.
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spelling nottingham-385132020-05-04T18:16:35Z https://eprints.nottingham.ac.uk/38513/ A sensitive 301V BSE serial PMCA assay Gough, Kevin C. Bishop, Keith Somerville, Robert A Hunter, Nora Maddison, Ben C. The prion strain 301V, is a mouse passaged form of bovine spongiform encephalopathy (BSE). It has been used as a model of BSE for more than 20 years, in particular in the investigation of tissue distribution of infectivity, the molecular phenotype and transmission properties of BSE, strain typing assays and prion inactivation studies. Most 301V experiments have required murine bioassay as a method for the quantitation of infectivity. To date this model strain has not been studied with the protein misfolding cyclic amplification assay (PMCA) which detects prion-associated PrPSc protein. The detection of BSE PrPSc by PMCA can be more sensitive than mouse bioassay and is carried out in a much shorter time frame of days as opposed to months/years. Here, we describe the development of a new highly sensitive and specific PMCA assay for murine 301V and assess the sensitivity of the assay in direct comparison with murine bioassay of the same material. This in vitro assay detected, in a few days, 301V at a brain dilution of at least 1x10-9, compared to bioassay of the same material in VM mice that could detect down to a 1x10-8 dilution and took >180 days. The 301V PMCA may therefore offer a faster and more sensitive alternative to live animal bioassay when studying the BSE agent in VM mice. F1000Research 2016-10-18 Article NonPeerReviewed Gough, Kevin C., Bishop, Keith, Somerville, Robert A, Hunter, Nora and Maddison, Ben C. (2016) A sensitive 301V BSE serial PMCA assay. F1000Research, 5 (2529). pp. 1-5. ISSN 2046-1402 https://f1000research.com/articles/5-2529/v1 doi:10.12688/f1000research.9735.1 doi:10.12688/f1000research.9735.1
spellingShingle Gough, Kevin C.
Bishop, Keith
Somerville, Robert A
Hunter, Nora
Maddison, Ben C.
A sensitive 301V BSE serial PMCA assay
title A sensitive 301V BSE serial PMCA assay
title_full A sensitive 301V BSE serial PMCA assay
title_fullStr A sensitive 301V BSE serial PMCA assay
title_full_unstemmed A sensitive 301V BSE serial PMCA assay
title_short A sensitive 301V BSE serial PMCA assay
title_sort sensitive 301v bse serial pmca assay
url https://eprints.nottingham.ac.uk/38513/
https://eprints.nottingham.ac.uk/38513/
https://eprints.nottingham.ac.uk/38513/