RT-QuIC and mass spectrometry approaches to prion strain detection and discrimination
- P.I. Byron Caughey, National Institutes of Health, Rocky Mountain Laboratories, USA
- Christina D. Orru, Research Fellow, Laboratory of Persistent Viral Diseases, NIH/NIAID Rocky Mountain Laboratories, USA
- Bradley R. Groveman, Laboratory of Persistent Viral Diseases, NIH/NIAID Rocky Mountain Laboratories, USA
- Roger Moore, Laboratory of Persistent Viral Diseases, NIH/NIAID Rocky Mountain Laboratories, USA
- Cristina Casalone, TSE surveillance center for the “Istituto Zooprofilattico Sperimentale del Piemonte Liguria e Valle d’Aosta” (IZSTO), Italy
- Cristiano Corona, TSE surveillance center for the IZSTO, Italy
- Maria Mazza, Genetica e Immunobiochimica, IZSTO, Italy
- Alessandra Favole, TSE surveillance center for the IZSTO, Italy
- Gianluigi Zanusso, University of Verona, Italy
2014: 18 months / Grant: 60 000 euros
Project: The goal of this project was to determine whether it was possible to discriminate prion
strains by RT-QuIC or RT-QuIC combined with mass spectrometric analysis, which had been shown
to differenciate animal prion strains. If it works, this could allow the distinction between type I and
II CJD, or BSE and BASE, which would be relevant for surveillance in animals and diagnosis in
Main results and related published data: By playing with different substrate sources of
recombinant PrP (rPrP: Syrian golden hamster rPrP23-231 or rPrP90-231, human rPrP23-231), and
chimeric hamster-sheep rPrP(23-137hamster+141-234ovine), it has been possible to discriminate the
L-BSE agent from the classical C-BSE agent in RT-QuIC assay. The amplification of PrPTSE from CBSE
could be improved and obtained only with a particular substrate (hamster-sheep rPrP), whereas
the amplification of PrPTSE from L-BSE could be obtained using multiple rPrP (hamster-sheep rPrP,
hamster rPrP 23-231 and 90-231, human 23-231). In addition, the limit of dilution with the hamstersheep
rPrP substrate is higher with L-BSE compared to C-BSE. Results reported recently completed
this study, since they showed that it was possible to discriminate the atypical H-BSE using multiple
rPrP subtrates (bank vole, ovine, chimeric hamster bovine… rPrP). Importantly bank vole rPrP also
appeared to be a virtually universalsubstrate for PrPTSE amplification using the RT-QuIC method for
many prion sources (vCJD, GSS, sCJD, several animal TSE and experimental models….).
- Detection and discrimination of classical and atypical L-type bovine spongiform
encephalopathy by real-time quaking-induced conversion. Orrú CD,Favole A, Corona C, Mazza
M, Manca M, Groveman BR, Hughson AG, Acutis PL, Caramelli M, Zanusso G, Casalone C, Caughey
B. J Clin Microbiol. 2015 Apr;53(4):1115-20. doi: 10.1128/JCM.02906-14. Epub 2015 Jan 21.
- Bank Vole Prion Protein As an Apparently Universal Substrate for RT-QuIC-Based
Detection and Discrimination of Prion Strains. Orrú CD, Groveman BR, Raymond LD, Hughson
AG, Nonno R, Zou W, Ghetti B, Gambetti P, Caughey B. PLoS Pathog. 2015 Jun 18;11(6):e1004983.
doi: 10.1371/journal.ppat.1004983. eCollection 2015.
- Detection of Atypical H-Type Bovine Spongiform Encephalopathy and Discrimination of
Bovine Prion Strains by Real-Time Quaking-Induced Conversion. Masujin K, Orrú CD,
Miyazawa K, Groveman BR, Raymond LD, Hughson AG, Caughey B. J Clin Microbiol. 2016
Mar;54(3):676-86. doi: 10.1128/JCM.02731-15. Epub 2016 Jan 6.
- Extended and direct evaluation of RT-QuIC assays for Creutzfeldt-Jakob disease diagnosis. Groveman BR, Orrú CD, Hughson AG, Bongianni M,Fiorini M, Imperiale D,Ladogana A, Pocchiari M,
Zanusso G, Caughey B. Ann Clin Transl Neurol. 2016 Dec 27;4(2):139-144. doi: 10.1002/acn3.378.