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. 2017 Dec 8;7:17241. doi: 10.1038/s41598-017-17090-x

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Detection of PrP^Sc in small volume of blood and removal of pre-cleaning step. To estimate the minimum amount of blood needed for detection, different volumes (20, 10, 5, 1 and 0.1 µL) of whole blood from five representative samples that were positive in the blinded study (3 B + LN + and 2 B-LN + ), were directly added to a 10% brain homogenate from cervid transgenic mice. As controls we used two samples from CWD negative animals. Samples were subjected to three sequential rounds of PMCA and PrP^Sc detected by Western blot using the PRC1 antibody. The figure shows the results of the third round of PMCA. As before, all samples were treated with PK, except the transgenic mice normal brain homogenate (NBH), used as migration control. Numbers in the right indicate the position of molecular weight markers.