In vitro amplification of ovine prions from scrapie-infected sheep from Great Britain reveals distinct patterns of propagation

BMC Veterinary Research

Research Article

Ben Charles Maddison¹  Jane Edwards²  Leigh Thorne²  Andrew Ramsay²  Otto Windl²  Thomas Holder²  Linda Ann Terry³  Maged Mohamed Taema⁴  Kevin Christopher Gough⁴ 

[1] ADAS UK, School of Veterinary Medicine and Science, The University of Nottingham, Sutton Bonington Campus, College Road, LE12 5RD, Sutton Bonington, Leicestershire, UK;Animal Health Veterinary Laboratories Agency (AHVLA), Woodham Lane, KT15 3NB, New Haw, Addlestone, Surrey, UK;Animal Health Veterinary Laboratories Agency (AHVLA), Woodham Lane, KT15 3NB, New Haw, Addlestone, Surrey, UK;Department of Immunology and Pathology, CMU, University of Geneva, 1 Rue Michel Servet, 1211, Geneva 4, Switzerland;School of Veterinary Medicine and Science, The University of Nottingham, Sutton Bonington Campus, College Road, LE12 5RD, Sutton Bonington, Leicestershire, UK;

关键词: Prions;    Transmissible spongiform encephalopathy;    Scrapie;    Sheep;    PMCA;   

DOI  :  10.1186/1746-6148-8-223

 received in 2012-07-06, accepted in 2012-10-30,  发布年份 2012

来源: Springer

【 摘 要 】

BackgroundProtein misfolding cyclic amplification (PMCA) is a method that facilitates the detection of prions from many sources of transmissible spongiform encephalopathy (TSE). Sheep scrapie represents a unique diversity of prion disease agents in a range of susceptible PRNP genotypes. In this study PMCA was assessed on a range of Great Britain (GB) sheep scrapie isolates to determine the applicability to veterinary diagnosis of ovine TSE.ResultsPrPSc amplification by protein misfolding cyclic amplification (PMCA) was assessed as a diagnostic tool for field cases of scrapie. The technique was initially applied to thirty-seven isolates of scrapie from diverse geographical locations around GB, and involved sheep of various breeds and PRNP genotypes. All samples were amplified in either VRQ and/or ARQ PrPC substrate. For PrPSc from sheep with at least one VRQ allele, all samples amplified efficiently in VRQ PrPC but only PrPSc from ARH/VRQ sheep amplified in both substrates. PrPSc from ARQ/ARQ sheep displayed two amplification patterns, one that amplified in both substrates and one that only amplified in ARQ PrPC. These amplification patterns were consistent for a further 14/15 flock/farm mates of these sheep. Furthermore experimental scrapie strains SSBP1, Dawson, CH1641 and MRI were analysed. SSBP1 and Dawson (from VRQ/VRQ sheep) amplified in VRQ but not ARQ substrate. MRI scrapie (from ARQ/ARQ sheep) nor CH1641 did not amplify in ARQ or VRQ substrate; these strains required an enhanced PMCA method incorporating polyadenylic acid (poly(A)) to achieve amplification.ConclusionsPrPsc from 52 classical scrapie GB field isolates amplified in VRQ or ARQ or both substrates and supports the use of PMCA as a rapid assay for the detection of a wide range of ovine classical scrapie infections involving multiple PRNP genotypes and scrapie strains.

【 授权许可】

Unknown   
© Thorne et al.; licensee BioMed Central Ltd. 2012. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.