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Updated: Jun 30, 2025
Real-time Quaking-induced Conversion Assay for Detection of CWD Prions in Fecal Material
Published on: September 29, 2017
Detection of prions in matching post-mortem skin and cerebrospinal fluid samples using second-generation real-time
Soňa Baranová1, Tibor Moško1, Magdalena Brůžová2
1Institute of Immunology and Microbiology, First Faculty of Medicine, Charles University, Prague, Czech Republic.
Abstract:
Real-time quaking-induced conversion assay (RT-QuIC) exploits templating activity of pathogenic prion protein for ultrasensitive detection of prions. We have utilized second generation RT-QuIC assay to analyze matching post-mortem cerebrospinal fluid and skin samples of 38 prion disease patients and of 30 deceased neurological controls. The analysis of cerebrospinal fluid samples led to 100% sensitivity and 100% specificity, but some samples had to be diluted before the analysis to alleviate the effect of present RT-QuIC inhibitors. The analysis of the corresponding skin samples provided 89.5% sensitivity and 100% specificity. The median seeding dose present in the skin was one order of magnitude higher than in the cerebrospinal fluid, despite the overall fluorescent signal of the skin samples was comparatively lower. Our data support the use of post-mortem cerebrospinal fluid for confirmation of prion disease diagnosis and encourage further studies of the potential of skin biopsy samples for intra-vitam prion diseases´ diagnostics.
Insights
The real-time quaking-induced conversion assay (RT-QuIC) offers ultrasensitive prion detection. Post-mortem cerebrospinal fluid achieved 100% accuracy for prion disease diagnosis, while skin samples showed high potential.
Area of Science:
- Neuroscience
- Biochemistry
- Medical Diagnostics
Background:
- Prion diseases are fatal neurodegenerative disorders.
- Accurate and early diagnosis is crucial for patient management.
- The real-time quaking-induced conversion assay (RT-QuIC) is a sensitive method for detecting misfolded prion proteins.
Purpose of the Study:
- To evaluate the diagnostic performance of the second-generation RT-QuIC assay using post-mortem cerebrospinal fluid (CSF) and skin samples.
- To compare the sensitivity and specificity of RT-QuIC in CSF versus skin for prion disease detection.
- To assess the potential of skin biopsy as a diagnostic tool for prion diseases.
Main Methods:
- Analysis of matching post-mortem CSF and skin samples from 38 prion disease patients and 30 controls using the second-generation RT-QuIC assay.
- CSF samples were sometimes diluted to mitigate RT-QuIC inhibitors.
- Quantification of prion seeding dose and fluorescent signal in both sample types.
Main Results:
- CSF analysis demonstrated 100% sensitivity and 100% specificity for prion disease diagnosis.
- Skin sample analysis yielded 89.5% sensitivity and 100% specificity.
- The median prion seeding dose in skin was higher than in CSF, despite lower overall fluorescence.
Conclusions:
- Post-mortem CSF is highly effective for confirming prion disease diagnosis via RT-QuIC.
- Skin samples show promising diagnostic potential for prion diseases, warranting further investigation for in vivo applications.
- RT-QuIC is a valuable tool for prion disease diagnostics, with potential applications in both CSF and skin.
