Journal Article

DZNE-2021-00192

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Optimization of the Real-Time Quaking-Induced Conversion Assay for Prion Disease Diagnosis.

Zerr, I. (First author)DZNE* ; Cramm, M.DZNE* ; Silva-Correia, S.DZNE* ; Zafar, S.DZNE* ; Villar-Piqué, A.DZNE* ; Llorens, F.DZNE* ; Schmitz, M. (Last author)DZNE*

2020
Frontiers Media Lausanne

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Please use a persistent id in citations: doi: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 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.

Keyword(s): Creutzfeldt-Jakob disease ; cerebrospinal fluid ; diagnostic test ; prion protein ; real-time quaking-induced conversion

Note: ISSN 2296-4185 not unique: **3 hits**.

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Contributing Institute(s):

1. Translational Studies and Biomarkers (AG Zerr)
2. Ext UMG Zerr (Ext UMG Zerr)

Research Program(s):

1. 344 - Clinical and Health Care Research (POF3-344) (POF3-344)

Appears in the scientific report 2020

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