Development of a sensitive real-time quaking-induced conversion

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 de...

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Bibliographic Details
Published inPloS one Vol. 18; no. 11; p. e0293845
Main Authors Thomas, Charlotte M, Salamat, M. Khalid F, de Wolf, Christopher, McCutcheon, Sandra, Blanco, A. Richard Alejo, Manson, Jean C, Hunter, Nora, Houston, E. Fiona
Format Journal Article
LanguageEnglish
Published Public Library of Science 02.11.2023
Subjects
Animal genetic engineering
Blood banks
Blood donors
Bovine spongiform encephalopathy
Brain
Care and treatment
Comparative analysis
Complications and side effects
Creutzfeldt-Jakob disease
Diagnosis
Disease transmission
Ethylenediaminetetraacetic acid
Evaluation
Ferric oxide
Infection
Iron compounds
Medical screening
Medical tests
Prion diseases
Prions
United Kingdom
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Summary: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 PrP.sup.Sc (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 PrP.sup.Sc 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 PrP.sup.Sc 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 PrP.sup.Sc 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.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0293845