Evaluation of Real time-QuIC and
enhanced QuIC assays
Evaluation of the Real Time QuIC and Enhanced QuIC assays for ante-mortem and
pre-symptomatic diagnosis of vCJD, sCJD, classical and atypical scrapie
- P.I. Christina Orru, University of Cagliari, Italy
- P.I. Sarah Vascellari, Department of Biomedical Sciences, Cagliari University, Italy
- Alessandra Pani, Department of Biomedical Sciences, Cagliari University, Italy
- Franco Cardone, Department of Cell Biology and Neurosciences, Istituto Superiore di Sanità, Italy
- Romolo Nonno DVM, Ph.D., Senior Scientist at the Department of Food Safety and Veterinary
Public Health, Istituto Superiore di Sanità, Rome, Italy.
- Byron Caughey,Laboratory of Persistent Viral Diseases, NIH/NIAID Rocky Mountain Laboratories, USA
2012: 1 year / Grant: 40 000 euros
Project: With this project, C. Orru aimed to use the RT-QuIC and eQuIC techniques which she helped
develop in Dr. B. Caughey’s laboratory, to develop an ante-mortem and pre-symptomatic diagnostic
test for human and ovine prion diseases.
Main results and related published data: Following the successful implementation of the
technique in Italy, the adaptation of the RT-QuIC to the detection of prion seeding activityin peripheral
samples allowed the consortium to develop protocols for prion detection based on RT-QuIC and
enhanced-QuIC. For example, the use of truncated substrates and SDS as well as a higher
temperature, improved the speed and sensitivity ofsCJD detection in CSFsamples. Recently C. Orru
also showed that bacterially expressed recombinant bank vole prion protein (residues 23-230) could
be used as a substrate for RT-QuIC detection of prions in several species (human, cattle, sheep,
cervids and rodents). In addition, discrimination between classical and atypical L-type bovine
spongiform encephalopathy,classical and atypical Nor98 scrapie in sheep, and sporadic and variant
Creutzfeldt-Jakob disease, could be performed using the bank vole substrate.
- Rapid and sensitive RT-QuIC detection of human Creutzfeldt-Jakob disease using
cerebrospinal fluid. Orrú CD, Groveman BR, Hughson AG, Zanusso G, Coulthart MB, Caughey B.
MBio. 2015 Jan 20;6(1). pii: e02451-14. doi: 10.1128/mBio.02451-14.
- 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.
- The real-time quaking-induced conversion assay for detection of human prion disease
and study of other protein misfolding diseases. Schmitz M, Cramm M,LlorensF, Müller-Cramm D, Collins S, Atarashi R, Satoh K, Orrù CD, Groveman BR, Zafar S, Schulz-Schaeffer WJ, Caughey B, Zerr I. Nat Protoc. 2016 Nov;11(11):2233-2242. doi: 10.1038/nprot.2016.120. Epub 2016 Oct 13.
- Factors That Improve RT-QuIC Detection of Prion Seeding Activity. Orrú CD, Hughson A, Groveman B, Campbell KJ, Anson KJ, Manca M, Kraus A, Caughey B. Viruses. 2016 May 23;8(5). pii: E140. doi: 10.3390/v8050140.
- 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.