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Published on: January 19, 2014
Differentiating ovine BSE from CH1641 scrapie by serial protein misfolding cyclic amplification
Maged M Taema1, Ben C Maddison, Leigh Thorne
1School of Veterinary Medicine and Science, The University of Nottingham, Sutton Bonington Campus, College Road, Sutton Bonington, Leicestershire LE12 5RD, UK.
Abstract:
Whilst ovine BSE displays distinct pathological characteristics to ovine CH1641-like scrapie upon passage in rodents, they have very similar molecular phenotypes. As such, the in vitro differentiation of these strains in routine surveillance programmes presents a significant diagnostic challenge. In this study, using serial protein-misfolding cyclic amplification (sPMCA), ovine BSE was readily amplified in vitro in brain substrates from sheep with V₁₃₆R₁₅₄Q₁₇₁/V₁₃₆R₁₅₄Q₁₇₁ or AHQ/AHQ PRNP genotypes. In contrast, the CH1641 strain was refractory to such amplification. This method allowed for complete and unequivocal differentiation of experimental BSE from CH1641 prion strains within an ovine host.
Insights
Differentiating ovine BSE from CH1641 scrapie is challenging due to similar molecular phenotypes. Serial protein-misfolding cyclic amplification (sPMCA) successfully distinguished these prion strains in vitro, aiding surveillance.
Area of Science:
- Veterinary Neurology
- Prion Disease Research
- Molecular Biology
Background:
- Ovine BSE and CH1641-like scrapie exhibit distinct pathological features but similar molecular phenotypes in rodents.
- This molecular similarity poses a significant diagnostic challenge for routine surveillance programs.
Purpose of the Study:
- To develop and validate an in vitro method for differentiating ovine BSE from CH1641 prion strains.
- To assess the efficacy of serial protein-misfolding cyclic amplification (sPMCA) for prion strain discrimination.
Main Methods:
- Utilized serial protein-misfolding cyclic amplification (sPMCA) to amplify prion strains in vitro.
- Employed brain substrates from sheep with specific PRNP genotypes (V136R154Q171/V136R154Q171 and AHQ/AHQ).
Main Results:
- Ovine BSE was readily amplified in vitro using sPMCA in the tested sheep brain substrates.
- The CH1641 prion strain proved refractory to amplification under the same sPMCA conditions.
- This differential amplification allowed for clear distinction between the two prion strains.
Conclusions:
- sPMCA provides a reliable method for the in vitro differentiation of ovine BSE and CH1641 prion strains.
- This technique can overcome diagnostic challenges posed by similar molecular phenotypes in routine surveillance.
- The study successfully differentiated experimental BSE from CH1641 prion strains within an ovine host.
