A sensitive 301 V 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 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 PrP protein. The detection of BSE PrP 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 assay detected, in a few in vitro days, 301V at a brain dilution of at least 1x10 , compared to bioassay of the same material in VM mice that could detect down to a 1x10 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. Ben C. Maddison ( ) Corresponding author: [email protected] Gough KC, Bishop K, Somerville RA How to cite this article: et al. A sensitive 301V BSE serial PMCA assay [version 1; referees: 1 2016, :2529 (doi: ) approved, 1 approved with reservations] F1000Research 5 10.12688/f1000research.9735.1 © 2016 Gough KC . This is an open access article distributed under the terms of the , which Copyright: et al Creative Commons Attribution Licence permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Data associated with the article are available under the terms of the (CC0 1.0 Public domain dedication). Creative Commons Zero "No rights reserved" data waiver This work was funded by DEFRA under project SE1433 (Robert Somerville) and by BBSRC Institute Strategic Grant Grant information: BB/J004332/1 (The Roslin Institute). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing interests: No competing interests were disclosed. 18 Oct 2016, :2529 (doi: ) First published: 5 10.12688/f1000research.9735.1 1 2 3 3