<it>In vitro</it> amplification of ovine prions from scrapie-infected sheep from Great Britain reveals distinct patterns of propagation
<p>Abstract</p> <p>Background</p> <p>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 agen...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2012-11-01
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| Series: | BMC Veterinary Research |
| Subjects: | |
| Online Access: | http://www.biomedcentral.com/1746-6148/8/223 |
| _version_ | 1818580346519683072 |
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| author | Thorne Leigh Holder Thomas Ramsay Andrew Edwards Jane Taema Maged Windl Otto Maddison Ben Charles Gough Kevin Christopher Terry Linda Ann |
| author_facet | Thorne Leigh Holder Thomas Ramsay Andrew Edwards Jane Taema Maged Windl Otto Maddison Ben Charles Gough Kevin Christopher Terry Linda Ann |
| author_sort | Thorne Leigh |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>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 <it>PRNP</it> 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.</p> <p>Results</p> <p>PrP<sup>Sc</sup> 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 <it>PRNP</it> genotypes. All samples were amplified in either VRQ and/or ARQ PrP<sup>C</sup> substrate. For PrP<sup>Sc</sup> from sheep with at least one VRQ allele, all samples amplified efficiently in VRQ PrP<sup>C</sup> but only PrP<sup>Sc</sup> from ARH/VRQ sheep amplified in both substrates. PrP<sup>Sc</sup> from ARQ/ARQ sheep displayed two amplification patterns, one that amplified in both substrates and one that only amplified in ARQ PrP<sup>C</sup>. 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.</p> <p>Conclusions</p> <p>PrP<sup>sc</sup> 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 <it>PRNP</it> genotypes and scrapie strains.</p> |
| first_indexed | 2024-12-16T07:16:08Z |
| format | Article |
| id | doaj.art-9bcaf064e5c34b1bb121f008dd353a56 |
| institution | Directory Open Access Journal |
| issn | 1746-6148 |
| language | English |
| last_indexed | 2024-12-16T07:16:08Z |
| publishDate | 2012-11-01 |
| publisher | BMC |
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| series | BMC Veterinary Research |
| spelling | doaj.art-9bcaf064e5c34b1bb121f008dd353a562022-12-21T22:39:46ZengBMCBMC Veterinary Research1746-61482012-11-018122310.1186/1746-6148-8-223<it>In vitro</it> amplification of ovine prions from scrapie-infected sheep from Great Britain reveals distinct patterns of propagationThorne LeighHolder ThomasRamsay AndrewEdwards JaneTaema MagedWindl OttoMaddison Ben CharlesGough Kevin ChristopherTerry Linda Ann<p>Abstract</p> <p>Background</p> <p>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 <it>PRNP</it> 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.</p> <p>Results</p> <p>PrP<sup>Sc</sup> 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 <it>PRNP</it> genotypes. All samples were amplified in either VRQ and/or ARQ PrP<sup>C</sup> substrate. For PrP<sup>Sc</sup> from sheep with at least one VRQ allele, all samples amplified efficiently in VRQ PrP<sup>C</sup> but only PrP<sup>Sc</sup> from ARH/VRQ sheep amplified in both substrates. PrP<sup>Sc</sup> from ARQ/ARQ sheep displayed two amplification patterns, one that amplified in both substrates and one that only amplified in ARQ PrP<sup>C</sup>. 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.</p> <p>Conclusions</p> <p>PrP<sup>sc</sup> 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 <it>PRNP</it> genotypes and scrapie strains.</p>http://www.biomedcentral.com/1746-6148/8/223PrionsTransmissible spongiform encephalopathyScrapieSheepPMCA |
| spellingShingle | Thorne Leigh Holder Thomas Ramsay Andrew Edwards Jane Taema Maged Windl Otto Maddison Ben Charles Gough Kevin Christopher Terry Linda Ann <it>In vitro</it> amplification of ovine prions from scrapie-infected sheep from Great Britain reveals distinct patterns of propagation BMC Veterinary Research Prions Transmissible spongiform encephalopathy Scrapie Sheep PMCA |
| title | <it>In vitro</it> amplification of ovine prions from scrapie-infected sheep from Great Britain reveals distinct patterns of propagation |
| title_full | <it>In vitro</it> amplification of ovine prions from scrapie-infected sheep from Great Britain reveals distinct patterns of propagation |
| title_fullStr | <it>In vitro</it> amplification of ovine prions from scrapie-infected sheep from Great Britain reveals distinct patterns of propagation |
| title_full_unstemmed | <it>In vitro</it> amplification of ovine prions from scrapie-infected sheep from Great Britain reveals distinct patterns of propagation |
| title_short | <it>In vitro</it> amplification of ovine prions from scrapie-infected sheep from Great Britain reveals distinct patterns of propagation |
| title_sort | it in vitro it amplification of ovine prions from scrapie infected sheep from great britain reveals distinct patterns of propagation |
| topic | Prions Transmissible spongiform encephalopathy Scrapie Sheep PMCA |
| url | http://www.biomedcentral.com/1746-6148/8/223 |
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