Journal
PRION
Volume 11, Issue 1, Pages 54-64Publisher
TAYLOR & FRANCIS INC
DOI: 10.1080/19336896.2016.1259051
Keywords
atypical prions; bovine spongiform encephalopathy; H-type BSE; PMCA
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Funding
- Biotechnology and Biological Sciences Research Council [BB/J014508/1]
- Biotechnology and Biological Sciences Research Council [1366329, 1270868] Funding Source: researchfish
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The 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 3 distinct forms, classical BSE (C-BSE) and 2 atypical BSE forms (L-BSE and H-BSE). Atypical forms are usually detected in 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 1x10(-12) g of brain material and was highly specific. Additionally, the product of serial protein misfolding cyclic amplification at all dilutions of seed analyzed 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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