OBJECTIVES: The core cerebrospinal fluid (CSF) biomarkers; total tau (tTau), phospho-tau (pTau), amyloid β 1-42 (Aβ 1-42), and the Aβ 1-42/Aβ 1-40 ratio have transformed Alzheimer's disease (AD) research and are today increasingly used in clinical routine laboratories as diagnostic tools. Fully automated immunoassay instruments with ready-to-use assay kits and calibrators has simplified their analysis and improved reproducibility of measurements. We evaluated the analytical performance of the fully automated immunoassay instrument LUMIPULSE G (Fujirebio) for measurement of the four core AD CSF biomarkers and determined cutpoints for AD diagnosis. METHODS: Comparison of the LUMIPULSE G assays was performed with the established INNOTEST ELISAs (Fujirebio) for hTau Ag, pTau 181, β-amyloid 1-42, and with V-PLEX Plus Aβ Peptide Panel 1 (6E10) (Meso Scale Discovery) for Aβ 1-42/Aβ 1-40, as well as with a LC-MS reference method for Aβ 1-42. Intra- and inter-laboratory reproducibility was evaluated for all assays. Clinical cutpoints for Aβ 1-42, tTau, and pTau was determined by analysis of three cohorts of clinically diagnosed patients, comprising 651 CSF samples. For the Aβ 1-42/Aβ 1-40 ratio, the cutpoint was determined by mixture model analysis of 2,782 CSF samples. RESULTS: The LUMIPULSE G assays showed strong correlation to all other immunoassays (r>0.93 for all assays). The repeatability (intra-laboratory) CVs ranged between 2.0 and 5.6%, with the highest variation observed for β-amyloid 1-40. The reproducibility (inter-laboratory) CVs ranged between 2.1 and 6.5%, with the highest variation observed for β-amyloid 1-42. The clinical cutpoints for AD were determined to be 409 ng/L for total tau, 50.2 ng/L for pTau 181, 526 ng/L for β-amyloid 1-42, and 0.072 for the Aβ 1-42/Aβ 1-40 ratio. CONCLUSIONS: Our results suggest that the LUMIPULSE G assays for the CSF AD biomarkers are fit for purpose in clinical laboratory practice. Further, they corroborate earlier presented reference limits for the biomarkers.

1st Department of Neurology Faculty of Medicine Masaryk University and St Anne's University Hospital Brno Czech Republic

Centro de Investigación Biomédica en Red Enfermedades Neurodegenerativas Madrid Spain

Clinical Neurochemistry Laboratory Sahlgrenska University Hospital Mölndal Sweden

Department of Laboratory Medicine Radboud University Medical Center Nijmegen The Netherlands

Department of Neurodegenerative Disease UCL Institute of Neurology London UK

Department of Neurology 2nd Medical Faculty Charles University Prague Czech Republic

Department of Neurology and Neurosurgery McGill University Research Centre for Studies in Aging Douglas Research Institute Le Centre intégré universitaire de santé et de services sociaux de l'Ouest de l'Île de Montréal Psychiatry and Pharmacology and Therapeutics McGill University Montreal QC Canada

Department of Neurology Memory Unit Hospital de la Santa Creu i Sant Pau Biomedical Research Institute Sant Pau Universitat Autònoma de Barcelona Barcelona Spain

Department of Neurology Radboud Alzheimer Centre Radboud University Medical Center Donders Institute for Brain Cognition and Behaviour Nijmegen The Netherlands

Department of Psychiatry and Neurochemistry Institute of Neuroscience and Physiology The Sahlgrenska Academy University of Gothenburg Gothenburg Sweden

Fujirebio Europe N 5 Ghent Belgium

International Clinical Research Center St Anne's University Hospital Brno Czech Republic

King's College London Institute of Psychiatry Psychology and Neuroscience Maurice Wohl Clinical Neuroscience Institute London UK

Laboratory of Clinical Neurochemistry Section of Neurology University of Perugia Perugia Italy

Montreal Neurological Institute Montreal QC Canada

Motol University Hospital Prague Czech Republic

NIHR Biomedical Research Centre for Mental Health and Biomedical Research Unit for Dementia at South London and Maudsley NHS Foundation London UK

Translational Neuroimaging Laboratory McGill Centre for Studies in Aging McGill University Montreal QC Canada

UK Dementia Research Institute at UCL London UK

Wallenberg Centre for Molecular and Translational Medicine University of Gothenburg Gothenburg Sweden

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