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Updated: Apr 9, 2026
Real-time Quaking-induced Conversion Assay for Detection of CWD Prions in Fecal Material
Published on: September 29, 2017
Bank Vole Prion Protein As an Apparently Universal Substrate for RT-QuIC-Based Detection and Discrimination of Prion
Christina D Orrú1, Bradley R Groveman1, Lynne D Raymond1
1Laboratory of Persistent Viral Diseases, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH), Hamilton, Montana, United States of America.
Abstract:
Prions propagate as multiple strains in a wide variety of mammalian species. The detection of all such strains by a single ultrasensitive assay such as Real Time Quaking-induced Conversion (RT-QuIC) would facilitate prion disease diagnosis, surveillance and research. Previous studies have shown that bank voles, and transgenic mice expressing bank vole prion protein, are susceptible to most, if not all, types of prions. Here we show that bacterially expressed recombinant bank vole prion protein (residues 23-230) is an effective substrate for the sensitive RT-QuIC detection of all of the different prion types that we have tested so far--a total of 28 from humans, cattle, sheep, cervids and rodents, including several that have previously been undetectable by RT-QuIC or Protein Misfolding Cyclic Amplification. Furthermore, comparison of the relative abilities of different prions to seed positive RT-QuIC reactions with bank vole and not other recombinant prion proteins allowed discrimination of prion strains such as classical and atypical L-type bovine spongiform encephalopathy, classical and atypical Nor98 scrapie in sheep, and sporadic and variant Creutzfeldt-Jakob disease in humans. Comparison of protease-resistant RT-QuIC conversion products also aided strain discrimination and suggested the existence of several distinct classes of prion templates among the many strains tested.
Insights
A novel ultrasensitive assay using bank vole prion protein enables the detection of all prion strains. This advancement in Real Time Quaking-induced Conversion (RT-QuIC) facilitates prion disease diagnosis and research.
Area of Science:
- Neuroscience
- Biochemistry
- Molecular Biology
Background:
- Prions exist as diverse strains across mammalian species.
- Accurate detection of all prion strains is crucial for diagnosis, surveillance, and research.
- Previous methods have limitations in detecting the full spectrum of prion strains.
Purpose of the Study:
- To develop a single ultrasensitive assay for detecting all prion strains.
- To evaluate the efficacy of recombinant bank vole prion protein as a substrate in RT-QuIC assays.
- To enable discrimination between different prion strains using RT-QuIC.
Main Methods:
- Utilized bacterially expressed recombinant bank vole prion protein (residues 23-230) as a substrate.
- Employed Real Time Quaking-induced Conversion (RT-QuIC) assay for prion detection.
- Compared prion seeding activity and conversion products across various prion types and recombinant prion proteins.
Main Results:
- The bank vole prion protein substrate successfully detected all 28 tested prion types from humans, cattle, sheep, cervids, and rodents.
- Several prion types previously undetectable by RT-QuIC or Protein Misfolding Cyclic Amplification were identified.
- RT-QuIC reactions with bank vole prion protein distinguished between specific prion strains, including BSE and CJD subtypes.
- Analysis of conversion products further aided strain discrimination and suggested distinct prion template classes.
Conclusions:
- Recombinant bank vole prion protein is a highly effective substrate for ultrasensitive RT-QuIC detection of diverse prion strains.
- This assay significantly advances the capability for prion disease diagnosis, surveillance, and research.
- RT-QuIC using bank vole prion protein allows for nuanced prion strain discrimination, aiding in understanding prion diversity and pathogenesis.
