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α-Synuclein Seeding Assay Using RT-QuIC.

Ayami Okuzumi1,2, Taku Hatano3, Takeshi Fukuhara3,4

  • 1Department of Neurology, Juntendo University School of Medicine, Tokyo, Japan. aokuzu@juntendo.ac.jp.

Methods in Molecular Biology (Clifton, N.J.)
|May 27, 2021
PubMed
Summary
This summary is machine-generated.

Real-time quaking-induced conversion (RT-QuIC) assays detect misfolded alpha-synuclein (αSyn) aggregates in synucleinopathies. These assays offer a sensitive method for diagnosing these neurodegenerative diseases.

Keywords:
BiomarkerBrainCerebrospinal fluidDiagnosisRT-QuICSeedStrainSynucleinopathyα-Synucleinβ-Sheet

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Area of Science:

  • Neuroscience
  • Biochemistry
  • Molecular Biology

Background:

  • Synucleinopathies, including Parkinson's disease, involve misfolded alpha-synuclein (αSyn) aggregation.
  • Distinct αSyn aggregate strains are proposed to cause disease-specific propagation and cell-to-cell spreading.
  • Seeded polymerization mechanisms are implicated in the growth of αSyn aggregates.

Purpose of the Study:

  • To detail protocols for alpha-synuclein (αSyn) real-time quaking-induced conversion (RT-QuIC) assays.
  • To highlight the application of αSyn RT-QuIC in diagnosing synucleinopathies.
  • To explore the utility of αSyn RT-QuIC in characterizing different αSyn strains.

Main Methods:

  • Utilizing the real-time quaking-induced conversion (RT-QuIC) assay mechanism, adapted from prion protein studies.
  • Incorporating thioflavin-T dye for real-time kinetic monitoring of αSyn aggregation.
  • Performing assays across multiple laboratories to assess performance.

Main Results:

  • The αSyn RT-QuIC assay demonstrates high sensitivity and specificity for detecting αSyn seeds.
  • Reported diagnostic specificity ranges from 82% to 100%, with sensitivity from 70% to 100%.
  • The assay enables sensitive detection of extremely low levels of seed aggregates.

Conclusions:

  • αSyn RT-QuIC assays provide a sensitive and specific diagnostic tool for synucleinopathies.
  • This method can differentiate between various αSyn strains implicated in different neurodegenerative diseases.
  • Detailed protocols are provided for the implementation and further study of αSyn RT-QuIC.