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A sensitive 301V BSE serial PMCA assay

Gough, Kevin C.; Bishop, Keith; Somerville, Robert A; Hunter, Nora; Maddison, Ben C.

Authors

KEVIN GOUGH kevin.gough@nottingham.ac.uk
Professor of Biochemistry and Pathology

Keith Bishop

Robert A Somerville

Nora Hunter

Ben C. Maddison



Abstract

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.

Journal Article Type Article
Publication Date Oct 18, 2016
Journal F1000Research
Electronic ISSN 2046-1402
Publisher F1000Research
Peer Reviewed Not Peer Reviewed
Volume 5
Issue 2529
APA6 Citation Gough, K. C., Bishop, K., Somerville, R. A., Hunter, N., & Maddison, B. C. (2016). A sensitive 301V BSE serial PMCA assay. F1000Research, 5(2529), https://doi.org/10.12688/f1000research.9735.1
DOI https://doi.org/10.12688/f1000research.9735.1
Publisher URL https://f1000research.com/articles/5-2529/v1
Copyright Statement Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0

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Copyright Statement
Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0





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