02141nas a2200253   4500000000100000000000100001008004100002260007000043653003700113653001600150653001100166653004400177653003800221100001600259700001400275700001300289700001300302700001500315700002400330245013000354490000800484520138100492022001401873       2018                            d  c06/2018bSchlütersche Verlagsgesellschaft mbH & Co. KGaHannover10abovine spongiform encephalopathy10ainfectivity10aPrPBSE10aProtein Misfolding Cyclic Amplification10areplacement of animal experiments1 aI Ackermann1 aJ Shawulu1 aM Keller1 aI Fatola1 aM Groschup1 aA Balkema-Buschmann00aExploring PMCA as a potential in-vitro alternative method to mouse bioassays for the highly sensitive detection of BSE prions0 v1313 aClassical bovine spongiform encephalopathy (C-BSE) belongs to the transmissible spongiform encephalopathies (TSE), which are also designated prion diseases since they are caused by the conversion of the host-encoded cellular prion protein PrPC to its pathological isoform PrPTSE. BSE carries a zoonotic potential as BSE prions cause variant Creutzfeldt-Jakob disease in humans. To date, C-BSE infectivity can only be detected by bioassay, e.g. highly sensitive bovine PrP transgenic mice (e.g. Tgbov XV mice). Recently, highly sensitive in-vitro prion seeding activity assays, such as the Protein Misfolding Cyclic Amplification (PMCA), have been developed, which work particularly well for the template-assisted prion conversion of scrapie prions, while a similarly efficient bovine C-BSE-prion amplification remained unavailable. In the here described study, we have therefore compared the analytical sensitivities of the transgenic Tgbov XV mouse bioassay and our C-BSE PMCA protocol by analysing serial dilutions of a BSE-positive bovine brainstem homogenate pool. As both methods were shown to possess comparable sensitivities, we propose the C-BSE PMCA as a potential in-vitro replacement method, allowing the reduction and refinement of mouse bioassays for the detection of cattle derived classical BSE prions by reducing them to only specific analytical applications.   a0005-9366