Rapid and ultra-sensitive quantitation of disease-associated α-synuclein seeds in brain and cerebrospinal fluid by αSyn RT-QuIC

Bradley R. Groveman, Christina D. Orrù, Andrew G. Hughson, Lynne D. Raymond, Gianluigi Zanusso, Bernardino Ghetti, Katrina J. Campbell, Jiri Safar, Douglas Galasko, Byron Caughey

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

The diagnosis and treatment of synucleinopathies such as Parkinson disease and dementia with Lewy bodies would be aided by the availability of assays for the pathogenic disease-associated forms of α-synuclein (αSynD) that are sufficiently sensitive, specific, and practical for analysis of accessible diagnostic specimens. Two recent αSynD seed amplification tests have provided the first prototypes for ultrasensitive and specific detection of αSynD in patients' cerebrospinal fluid. These prototypic assays require 5-13days to perform. Here, we describe an improved α-synuclein real time quaking-induced conversion (αSyn RT-QuIC) assay that has similar sensitivity and specificity to the prior assays, but can be performed in 1-2days with quantitation. Blinded analysis of cerebrospinal fluid from 29 synucleinopathy cases [12 Parkinson's and 17 dementia with Lewy bodies] and 31 non-synucleinopathy controls, including 16 Alzheimer's cases, yielded 93% diagnostic sensitivity and 100% specificity for this test so far. End-point dilution analyses allowed quantitation of relative amounts of αSynD seeding activity in cerebrospinal fluid samples, and detection in as little as 0.2μL. These results confirm that αSynD seeding activity is present in cerebrospinal fluid. We also demonstrate that it can be rapidly detected, and quantitated, even in early symptomatic stages of synucleinopathy.

Original languageEnglish (US)
Article number7
JournalJournal of Biological Research (Greece)
Volume25
Issue number1
DOIs
StatePublished - Feb 8 2018

Fingerprint

Synucleins
Cerebrospinal fluid
cerebrospinal fluid
Cerebrospinal Fluid
Seed
Assays
Brain
Seeds
brain
Lewy Body Disease
dementia
assays
seeds
sowing
diagnostic sensitivity
Sensitivity and Specificity
Parkinson disease
prototypes
Dilution
Amplification

Keywords

  • Alzheimer
  • Amplification
  • Cerebrospinal fluid
  • Diagnosis
  • Lewy body
  • Parkinson
  • PMCA
  • Prion
  • RT-QuIC
  • Synuclein

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Rapid and ultra-sensitive quantitation of disease-associated α-synuclein seeds in brain and cerebrospinal fluid by αSyn RT-QuIC. / Groveman, Bradley R.; Orrù, Christina D.; Hughson, Andrew G.; Raymond, Lynne D.; Zanusso, Gianluigi; Ghetti, Bernardino; Campbell, Katrina J.; Safar, Jiri; Galasko, Douglas; Caughey, Byron.

In: Journal of Biological Research (Greece), Vol. 25, No. 1, 7, 08.02.2018.

Research output: Contribution to journalArticle

Groveman, Bradley R. ; Orrù, Christina D. ; Hughson, Andrew G. ; Raymond, Lynne D. ; Zanusso, Gianluigi ; Ghetti, Bernardino ; Campbell, Katrina J. ; Safar, Jiri ; Galasko, Douglas ; Caughey, Byron. / Rapid and ultra-sensitive quantitation of disease-associated α-synuclein seeds in brain and cerebrospinal fluid by αSyn RT-QuIC. In: Journal of Biological Research (Greece). 2018 ; Vol. 25, No. 1.
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