ArticleClinical chemistry and laboratory medicine2024
Improving protocols for α-synuclein seed amplification assays: analysis of preanalytical and analytical variables and identification of candidate parameters for seed quantification.
Article in Clinical chemistry and laboratory medicine, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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Who cites it
25 citing papers in PubMed.
- Inter-assay and Inter-site Performance of Alpha-Synuclein Seed Amplification Assays in Synucleinopathies: A Multicenter 2 × 2 Protocol Comparison.Neurology and therapy · 2026Article
- Characterization of patients with isolated REM sleep behavior disorder and cerebrospinal fluid α-synuclein negativity.NPJ Parkinson's disease · 2026Article
- A Systematic Comparison of Alpha-Synuclein Seed Amplification Assays for Increasing Reproducibility.Annals of clinical and translational neurology · 2026Review
- Towards biomarker-based diagnosis of Parkinson disease.Nature reviews. Neurology · 2026Review
- When Proteins Go MAD-Misfolded, Amplified, Detected: Advances in α-Synuclein Pathophysiology and RT-QuIC Detection.Molecular neurobiology · 2026Review
- Quantification of cerebrospinal fluid α-synuclein seeds by endpoint dilution seed amplification assay in Parkinson's disease.NPJ Parkinson's disease · 2025Article
- Quantitative measurements of α-synuclein seeds in CSF inform diagnosis of synucleinopathies.Journal of Parkinson's disease · 2025Article
- Dermal Alpha-Synuclein Aggregation in Seed Amplification Assays for Parkinson's Disease Subtype Differentiation.European journal of neurology · 2025Article
- α-Synuclein purification significantly impacts seed amplification assay performance and consistency.Acta neuropathologica communications · 2025Article
- Cognitive function correlates with gastric alpha-synuclein seeding activity in early Parkinson's disease.NPJ Parkinson's disease · 2025Article
- Comparison of Two α-Synuclein Seed Amplification Assays for Discrimination of Parkinson Disease and Atypical Parkinsonism.Movement disorders : official journal of the Movement Disorder Society · 2025Article
- Fluid biomarkers in atypical Parkinsonism: current state and future perspectives.Journal of neural transmission (Vienna, Austria : 1996) · 2025Review
- Alpha-synuclein seeding amplification assays in Lewy body dementia: a brief review.Molecular neurodegeneration · 2025Review
- Quantification of Lewy body pathology by cerebrospinal fluid endpoint dilution RT-QuIC in a neuropathological autopsy cohort of clinically heterogeneous participants.Acta neuropathologica · 2025Article
- Alpha-synuclein seed amplification assay longitudinal outcomes in Lewy body disease spectrum.Brain : a journal of neurology · 2025Article
- Advancing Parkinson's diagnosis: seed amplification assay for α-synuclein detection in minimally invasive samples.Molecular and cellular biochemistry · 2025Review
- Seeding amplification assay: Limitations and insights for enhanced clinical and research applications.Journal of Parkinson's disease · 2025Review
- Head-to-head comparison of four cerebrospinal fluid and three plasma neurofilament light chain assays in Parkinsonism.NPJ Parkinson's disease · 2025Article
- A quantitative Lewy-fold-specific alpha-synuclein seed amplification assay as a progression marker for Parkinson's disease.Acta neuropathologica · 2025Article
- Refining α-synuclein seed amplification assays to distinguish Parkinson's disease from multiple system atrophy.Translational neurodegeneration · 2025Article
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Abstract
objectivesThe effect of preanalytical and analytical factors on the α-synuclein (α-syn) seed amplification assay's (SAA) performance has not been fully explored. Similarly, there is limited knowledge about the most suitable assay protocol and kinetic parameters for misfolded α-syn seed quantification.
methodsWe studied the effect of centrifugation, repeated freeze-thaw cycles (up to seven), delayed freezing, detergent addition, and blood contamination on the performance of the cerebrospinal fluid (CSF) α-syn SAA real-time quaking-induced conversion (RT-QuIC). Moreover, we analysed the inter- and intra-plate variability, the recombinant protein batch effect, and the RT-QuIC parameters' variability when multiple samples were run in controlled conditions. Finally, we evaluated the assay potential of quantifying α-syn seed by assessing kinetic curves in serial CSF dilutions.
resultsAmong tested preanalytical variables, a ≥0.01 % blood contamination and adding detergents significantly affected the RT-QuIC kinetic parameters and the number of positive replicates. Increasing the number of replicates improved result reproducibility. The number of positive replicates in serially diluted CSF samples improved discrimination between samples with high and low seeding activity, and the time to threshold (LAG) was the most reliable kinetic parameter in multiple experiment settings.
conclusionsPreanalytical variables affecting α-syn RT-QuIC performance are limited to blood contamination and detergent addition. The number of positive replicates and the LAG are the most reliable variables for quantifying α-syn seeding activity. Their consistent measurement in serial dilution experiments, especially when associated with an increased number of sample replicates, will help to develop the α-syn RT-QuIC assay further into a quantitative test.
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