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Updated: Jun 27, 2026
Assessing Transmissible Spongiform Encephalopathy Species Barriers with an In Vitro Prion Protein Conversion Assay
Published on: March 10, 2015
Development of a sensitive real-time quaking-induced conversion (RT-QuIC) assay for application in prion-infected
Charlotte M Thomas1, M Khalid F Salamat1, Christopher de Wolf1
1The Roslin Institute, Royal (Dick) School of Veterinary Studies, University of Edinburgh, Easter Bush Campus, Midlothian, United Kingdom.
Abstract:
Efforts to prevent human-to-human transmission of variant Creutzfeldt-Jakob disease (vCJD) by contaminated blood would be aided by the development of a sensitive diagnostic test that could be routinely used to screen blood donations. As blood samples from vCJD patients are extremely rare, here we describe the optimisation of real-time quaking-induced conversion (RT-QuIC) for detection of PrPSc (misfolded prion protein, a marker of prion infection) in blood samples from an established large animal model of vCJD, sheep experimentally infected with bovine spongiform encephalopathy (BSE). Comparative endpoint titration experiments with RT-QuIC, miniaturized bead protein misfolding cyclic amplification (mb-PMCA) and intracerebral inoculation of a transgenic mouse line expressing sheep PrP (tgOvARQ), demonstrated highly sensitive detection of PrPSc by RT-QuIC in a reference sheep brain homogenate. Upon addition of a capture step with iron oxide beads, the RT-QuIC assay was able to detect PrPSc in whole blood samples from BSE-infected sheep up to two years before disease onset. Both RT-QuIC and mb-PMCA also demonstrated sensitive detection of PrPSc in a reference vCJD-infected human brain homogenate, suggesting that either assay may be suitable for application to human blood samples. Our results support the further development and evaluation of RT-QuIC as a diagnostic or screening test for vCJD.
Insights
A new diagnostic test, real-time quaking-induced conversion (RT-QuIC), shows promise for detecting prion disease (vCJD) in blood. This sensitive method could aid in screening blood donations to prevent disease transmission.
Area of Science:
- Neuroscience
- Biochemistry
- Veterinary Medicine
Background:
- Variant Creutzfeldt-Jakob disease (vCJD) poses a risk of transmission through contaminated blood.
- Sensitive diagnostic tests are crucial for screening blood donations to prevent vCJD spread.
- Prion protein (PrPSc) is a marker for prion infection.
Purpose of the Study:
- To optimize real-time quaking-induced conversion (RT-QuIC) for detecting PrPSc in blood.
- To evaluate RT-QuIC's sensitivity compared to other methods in an animal model.
- To assess the potential of RT-QuIC for screening human blood donations for vCJD.
Main Methods:
- Optimization of RT-QuIC assay for PrPSc detection in blood.
- Comparative analysis of RT-QuIC, mb-PMCA, and transgenic mouse inoculation.
- Development of a capture step using iron oxide beads for enhanced PrPSc detection.
Main Results:
- RT-QuIC demonstrated highly sensitive PrPSc detection in sheep brain homogenates.
- RT-QuIC detected PrPSc in whole blood of BSE-infected sheep up to two years pre-disease onset.
- Both RT-QuIC and mb-PMCA detected PrPSc in vCJD-infected human brain homogenates.
Conclusions:
- RT-QuIC is a highly sensitive method for detecting PrPSc in blood samples from an animal model of vCJD.
- The assay shows potential for early detection of PrPSc in blood, aiding transfusion safety.
- Further development and evaluation of RT-QuIC are supported for vCJD diagnostic and screening applications.
