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Short-interval variability and test-retest reliability of Alzheimer’s disease plasma biomarkers in a clinically heterogenous cohort

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Beament, Millie, Wiltshire, Katharine, Belder, Christopher R S, Abel, Emily, Hart, Melanie, Heslegrave, Amanda J, Lunn, Michael P, Rosa-Grilo, Miguel, Swann, Owen, Schott, Jonathan M, Zetterberg, Henrik, Fox, Nick C, Keshavan, Ashvini·2026·Alzheimer's Research & Therapy
DOI 10.1186/s13195-026-02180-8PMC PMC13711881
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Abstract

Background: Assessment of short-term within-subject variability and test-retest reliability of Alzheimer’s disease (AD)-related plasma biomarkers is important for real-world clinical implementation and interpretation in research studies. In this study we aimed to assess short-term variability and reliability of core AD plasma biomarkers, proposed ratios, and evaluate a broader proteomic panel. Methods: Two serial plasma samples were collected from each of the 92 participants enrolled in observational research studies over intervals ranging from 30 minutes to three months. Test-retest reliability and within-subject variability were assessed for 131 biomarkers measured on three immunoassays (Lumipulse G plasma p-tau217; Fujirebio, NULISAseq CNS Disease Panel and NULISAqpcr AD 5-Plex; Alamar Biosciences). We subsequently evaluated the stability of a dual cut-point approach for AD probability diagnostic categorisation and examined whether within-subject variability was associated with inter-sample interval (time between serial blood sampling). Results: All core AD plasma biomarkers assessed in this study (Aβ42, BD-p-tau181, BD-p-tau217, BD-p-tau231, p-tau181, p-tau217, p-tau231, GFAP and NfL) demonstrated strong correlations (r ≥ 0.79) and high reliability (ICC ≥ 0.84). Within-subject variability was moderate to low for core AD plasma biomarkers. Using a validated dual cut-point approach for plasma p-tau217, most individuals (92%) remained within the same diagnostic category (high/indeterminate/low) across both timepoints, and no individual moved across two boundaries (i.e. from high to low AD probability categorisation or vice versa). For the majority of analytes, within-subject variability was not associated with inter-sample interval after adjustment for age and sex, however, the core AD plasma biomarker, NfL did have a significant association. Conclusions: While core AD plasma biomarkers had limited variability and high reliability over short-intervals, other central nervous system (CNS)-relevant biomarkers showed greater variability, which may be relevant for interpretation in association studies and clinical applications.

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Metadata

Authors
Beament, M., Wiltshire, K., Belder, C. R. S., Abel, E., Hart, M., Heslegrave, A. J., Lunn, M. P., Rosa-Grilo, M., Swann, O., Schott, J. M., Zetterberg, H., Fox, N. C., & Keshavan, A.
Affiliations
  • UK Dementia Research Institute (GB)
  • Royal Adelaide Hospital (AU)
  • National Hospital for Neurology and Neurosurgery (GB)
  • University of Wisconsin–Madison (US)
Corresponding author
Beament, Millie
ORCID
Venue
Alzheimer's Research & Therapy
Publisher
Springer Science and Business Media LLC
Published
2026-10-27
Volume
18
Issue
1
ISSN
1758-9193 (ISSN-L)
DOI
10.1186/s13195-026-02180-8
PMC
PMC13711881
OpenAlex
W7223785278
Cited by
0
References
2
FWCI
0.00
Type
journal article
Language
en
Open access
Licence
Locations
  • Alzheimer s Research & Therapy (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • PubMed Central (publishedVersion, cc-by)
  • Figshare (cc-by)
Funders
  • Hospital Research Foundation
  • Alzheimer's Society
  • National Institute for Health and Care Research
  • British Heart Foundation (RE/24/130013)
  • University College London
  • Rosetrees Trust
  • University College London Hospitals NHS Foundation Trust
  • UK Dementia Research Institute
  • Medical Research Council
  • National Health and Medical Research Council (2049937)
  • UCLH Biomedical Research Centre
Topics
  • Health Sciences › Medicine › Psychiatry and Mental health › Dementia and Cognitive Impairment Research
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Alzheimer's disease research and treatments
  • Health Sciences › Health Professions › Medical Laboratory Technology › Clinical Laboratory Practices and Quality Control
Keywords
Alzheimer's disease, plasma biomarkers, test-retest reliability, within-person variability, p-tau217, phosphorylated tau, amyloid-beta 42, neurofilament light chain, glial fibrillary acidic protein
MeSH terms
Aged, Aged, 80 and over, Alzheimer Disease, Female, Glial Fibrillary Acidic Protein, Humans, Male, Peptide Fragments, Reproducibility of Results, Cohort Studies, Biomarkers, Amyloid beta-Peptides, tau Proteins

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