Development of an Automated Capillary Immunoassay to Detect Prion Glycotypes in Creutzfeldt-Jakob Disease

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DOI

https://doi.org/10.1016/j.labinv.2022.100029

Language of the publication
English
Date
2023-03
Type
Article
Author(s)
  • Myskiw, Jennifer
  • Lamoureux, Lise
  • Peterson, Anne
  • Knox, David
  • Jansen, Gerald H.
  • Coulthart, Michael B.
  • Booth, Stephanie A.
Publisher
Nature

Abstract

Creutzfeldt-Jakob disease (CJD) comprises a group of transmissible neurodegenerative diseases with vast phenotypic diversity. Sporadic CJD heterogeneity is predominantly influenced by the genotype at codon 129 of the prion-encoding gene and the molecular weight of PrPSc fragments after protease digestion, resulting in a classification of 6 subtypes of CJD (MM1, MM2, MV1, MV2, VV1, and VV2). The majority of cases with CJD can be distinguished using this classification system. However, a number of reported CJD cases are phenotypically unique from others within their same subtype, such as variably protease-sensitive prionopathies, or exist as a mixture of subtypes within the same patient. Western blotting of brain tissue, along with the genotyping of codon 129 of the prion-encoding gene, is considered the "gold standard" for the biochemical characterization of CJD. Western blotting requires a significant amount of prion protein for detection, is labor-intensive, and is also associated with high interassay variability. In addition to these limitations, a growing body of research suggests that unique subtypes of CJD are often undetected or misdiagnosed using standard diagnostic western blotting protocols. Consequently, we successfully optimized and developed a capillary-based western assay using the JESS Simple Western (ProteinSimple) to detect and characterize prion proteins from patients with CJD. We found that this novel assay consistently differentiated CJD type 1 and type 2 cases with a limit of detection 10 to 100× higher than traditional western blotting. Cases with CJD in which type 1 and type 2 coexist within the same brain region can be detected using type 1-specific and type 2-specific antibodies, and we found that there was remarkable specificity for the detection of cases with variably protease-sensitive prionopathy. The assay presented displays outstanding sensitivity, allowing for the preservation of valuable samples and enhancing current detection methods.

Subject

  • Health

Keywords

  • Creutzfeldt-Jakob disease,
  • Creutzcapillary electrophoresis,
  • Prion

Rights

Open Government Licence - Canada

Peer review

Yes

Open access level

Green

Identifiers

ISSN
1530-0307

Article

Journal title
Laboratory Investigation
Journal volume
103
Journal issue
3

Citation(s)

Myskiw J, Lamoureux L, Peterson A, Knox D, Jansen GH, Coulthart MB, Booth SA. Development of an Automated Capillary Immunoassay to Detect Prion Glycotypes in Creutzfeldt-Jakob Disease. Lab Invest. 2023 Mar;103(3):100029. doi: 10.1016/j.labinv.2022.100029

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Communicable diseases

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