Optimization of the Real-Time Quaking-Induced Conversion Assay for Prion Disease Diagnosis
2020 | journal article. A publication with affiliation to the University of Göttingen.
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Optimization of the Real-Time Quaking-Induced Conversion Assay for Prion Disease Diagnosis
Zerr, I. ; Cramm, M. ; da Silva Correia, S. M.; Zafar, S. ; Villar-Piqué, A. ; Llorens, F. & Schmitz, M. (2020)
Frontiers in Bioengineering and Biotechnology, 8. DOI: https://doi.org/10.3389/fbioe.2020.586890
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- Authors
- Zerr, Inga ; Cramm, Maria ; da Silva Correia, Susana Margarida; Zafar, Saima ; Villar-Piqué, Anna ; Llorens, Franc ; Schmitz, Matthias
- Abstract
- The real-time quaking-induced conversion (RT-QuIC) assay is a highly reproducible and robust methodology exhibiting an excellent pre-mortem diagnostic accuracy for prion diseases. However, the protocols might be time-consuming and improvement of the detection technology is needed. In the present study, we investigated the influence of a pre-analytical cerebrospinal fluid (CSF) treatment with proteinase K (PK) on the kinetic of the RT-QuIC signal response. For this purpose, we added PK at different concentrations in RT-QuIC reactions seeded with Creutzfeldt–Jakob disease (sCJD) CSF. We observed that a mild pre-analytical PK treatment of CSF samples resulted in an increased seeding efficiency of the RT-QuIC reaction. Quantitative seeding parameters, such as a higher area under the curve (AUC) value or a shorter lag phase indicated a higher conversion efficiency after treatment. The diagnostic accuracy resulting from 2 μg/ml PK treatment was analyzed in a retrospective study, where we obtained a sensitivity of 89%. Additionally, we analyzed the agreement with the previously established standard RT-QuIC protocol without PK treatment in a prospective study. Here, we found an overall agreement of 94% to 96%. A Cohen’s kappa of 0.9036 (95% CI: 0.8114–0.9958) indicates an almost perfect agreement between both protocols. In conclusion, the outcome of our study can be used for a further optimization of the RT-QuIC assay in particular for a reduction of the testing time.
- Issue Date
- 2020
- Publisher
- Frontiers Media S.A.
- Journal
- Frontiers in Bioengineering and Biotechnology
- eISSN
- 2296-4185
- Language
- English
- Sponsor
- Open-Access-Publikationsfonds 2020