In vitro amplification of H-type atypical bovine spongiform encephalopathy by protein misfolding cyclic amplificationTools O'Connor, M.J., Bishop, Keith, Workman, Robert G., Maddison, Ben C. and Gough, Kevin C. (2017) In vitro amplification of H-type atypical bovine spongiform encephalopathy by protein misfolding cyclic amplification. Prion, 11 (1). pp. 54-64. ISSN 1933-690X Full text not available from this repository.AbstractThe in vitro amplification of prions by serial protein misfolding cyclic amplification has been shown to detect PrPSc to levels at least as sensitive as rodent bioassay but in a fraction of the time. Bovine spongiform encephalopathy is a zoonotic prion disease in cattle and has been shown to occur in three distinct forms, classical BSE (C-BSE) and two atypical BSE forms (L-BSE and H-BSE). Atypical forms are usually detectedin asymptomatic, older cattle and are suggested to be spontaneous forms of the disease. Here, we show the development of a serial protein misfolding cyclic amplification method for the detection of H-BSE. The assay could detect PrPSc from 3 distinct experimental isolates of H-BSE, could detect PrPSc in as little as 1 x 10-12 g of brain material and was highly specific. Additionally, the product of serial protein misfolding cyclic amplification at all dilutions of seed analysed could be readily distinguished from L-BSE, which did not amplify, and C-BSE, which had PrPSc with distinct protease K-resistance and protease K-resistant PrPSc molecular weights.
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