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2020-11-19

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cc by (c) Zerr et al., 2020
Please use this identifier to cite or link to this item: https://hdl.handle.net/2445/173811

Optimization of the Real-Time Quaking-Induced Conversion Assay for Prion Disease Diagnosis

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https://doi.org/10.3389/fbioe.2020.586890

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 mu g/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.

Subject

Malaltia de Creutzfeldt-Jakob, Prions

Subject (English)

Creutzfeldt-Jakob disease, Prions

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Articles publicats en revistes (Institut d'lnvestigació Biomèdica de Bellvitge (IDIBELL))

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ZERR, Inga, et al. Optimization of the Real-Time Quaking-Induced Conversion Assay for Prion Disease Diagnosis. Frontiers in Bioengineering And Biotechnology. 2020. Vol. 8. [consulted: 8 of August of 2026]. Available at: https://hdl.handle.net/2445/173811

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