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

BMC Veterinary Research

Linda Ann Terry¹  Kevin Christopher Gough²  Ben Charles Maddison⁴  Otto Windl³  Maged Mohamed Taema²  Jane Edwards³  Andrew Ramsay³  Thomas Holder³  Leigh Thorne³ 

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

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

Others  :  1119664
DOI  :  10.1186/1746-6148-8-223

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

【 摘 要 】

Background

Protein 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.

Results

PrP^Sc 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 PrP^C substrate. For PrP^Sc from sheep with at least one VRQ allele, all samples amplified efficiently in VRQ PrP^C but only PrP^Sc from ARH/VRQ sheep amplified in both substrates. PrP^Sc from ARQ/ARQ sheep displayed two amplification patterns, one that amplified in both substrates and one that only amplified in ARQ PrP^C. 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.

Conclusions

PrP^sc 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】

   
2012 Thorne et al.; licensee BioMed Central Ltd.

【 图 表 】

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