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Development and multi-center validation of a fully automated digital immunoassay for neurofilament light chain: toward a clinical blood test for neuronal injury

  • David Wilson , Dandan Chan , Lei Chang , Robert Mathis , Inge Verberk , Xavier Montalban , Manuel Comabella , Nicolas Fissolo , Bibi Bielekova , Ruturaj Masvekar , Tanuja Chitnis , Tjalf Ziemssen , Katja Akgün , Kaj Blennow , Henrik Zetterberg , Wolfgang Brück , Gavin Giovannoni , Sharmilee Gnanapavan , Stefan Bittner , Frauke Zipp , Giancarlo Comi , Roberto Furlan , Sylvain Lehmann , Simon Thebault , Mark Freedman , Amit Bar-Or , Marty Kramer , Markus Otto , Steffen Halbgebauer , Kevin Hrusovsky , Tatiana Plavina , Michael Khalil , Fredrik Piehl , Heinz Wiendl , Ludwig Kappos , Aleksandra Maceski , Eline Willemse , David Leppert , Charlotte Teunissen and Jens Kuhle EMAIL logo
Published/Copyright: September 13, 2023
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Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 62 Issue 2

Abstract

Objectives

Neurofilament light chain (NfL) has emerged as a promising biomarker for detecting and monitoring axonal injury. Until recently, NfL could only be reliably measured in cerebrospinal fluid, but digital single molecule array (Simoa) technology has enabled its precise measurement in blood samples where it is typically 50–100 times less abundant. We report development and multi-center validation of a novel fully automated digital immunoassay for NfL in serum for informing axonal injury status.

Methods

A 45-min immunoassay for serum NfL was developed for use on an automated digital analyzer based on Simoa technology. The analytical performance (sensitivity, precision, reproducibility, linearity, sample type) was characterized and then cross validated across 17 laboratories in 10 countries. Analytical performance for clinical NfL measurement was examined in individual patients with relapsing remitting multiple sclerosis (RRMS) after 3 months of disease modifying treatment (DMT) with fingolimod.

Results

The assay exhibited a lower limit of detection (LLoD) of 0.05 ng/L, a lower limit of quantification (LLoQ) of 0.8 ng/L, and between-laboratory imprecision <10 % across 17 validation sites. All tested samples had measurable NfL concentrations well above the LLoQ. In matched pre–post treatment samples, decreases in NfL were observed in 26/29 RRMS patients three months after DMT start, with significant decreases detected in a majority of patients.

Conclusions

The sensitivity characteristics and reproducible performance across laboratories combined with full automation make this assay suitable for clinical use for NfL assessment, monitoring in individual patients, and cross-comparisons of results across multiple sites.

Keywords: digital immunoassay; neuronal injury; multiple sclerosis; validation
Corresponding author: Jens Kuhle, MD, PhD, Multiple Sclerosis Centre and Research Center for Clinical Neuroimmunology and Neuroscience Basel (RC2NB), Departments of Head, Spine and Neuromedicine, Biomedicine and Clinical Research, University Hospital Basel and University of Basel, Basel, Switzerland, Phone: +41 61 265 2525, E-mail:

  1. Research ethics: The local Institutional Review Board deemed the study exempt from review.

  2. Informed consent: Informed consent was obtained from all individuals included in this study.

  3. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.

  4. Competing interests: David Wilson is an employee at Quanterix. Dandan Chan was an employee at Quanterix when the work was completed. Lei Chang was an employee at Quanterix when the work was completed. Robert Mathis was an employee at Quanterix when the work was completed. Inge Verberk states no conflict of interest. Xavier Montalban states no conflict of interest. Manuel Comabella states no conflict of interest. Nicolas Fissolo states no conflict of interest. Bibi Bielekova states no conflict of interest. Ruturaj Masvekar states no conflict of interest. Tanuja Chitnis states no conflict of interest. Tjalf Ziemssen states no conflict of interest. Katja Akgün states no conflict of interest. Kaj Blennow states no conflict of interest. Henrik Zetterberg states no conflict of interest. Wolfgang Brück states no conflict of interest. Gavin Giovannoni states no conflict of interest. Sharmilee Gnanapavan states no conflict of interest. Stefan Bittner states no conflict of interest. Frauke Zipp states no conflict of interest. Giancarlo Comi states no conflict of interest. Roberto Furlan states no conflict of interest. Sylvain Lehmann states no conflict of interest. Simon Thebault states no conflict of interest. Mark Freedman has consulted for Quanterix and received consultant fees. Amit Bar-Or states no conflict of interest. Marty Kramer states no conflict of interest. Markus Otto states no conflict of interest. Steffen Halbgebauer states no conflict of interest. Kevin Hrusovsky was an employee at Quanterix when the work was completed. Tatiana Plavina states no conflict of interest. Michael Khalil states no conflict of interest. Fredrik Piehl states no conflict of interest. Heinz Wiendl states no conflict of interest. Ludwig Kappos states no conflict of interest. Aleksandra Maceski states no conflict of interest. Eline Willemse states no conflict of interest. David Leppert states no conflict of interest. Charlotte Teunissen has a collaboration contract with Quanterix. Jens Kuhle has consulted for Quanterix and received consultant fees.

  5. Research funding: None declared.

  6. Data availability: The data that support the findings of this study are available from the corresponding author, JK, upon reasonable request.

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Supplementary Material

This article contains supplementary material (https://doi.org/10.1515/cclm-2023-0518).

Received: 2023-05-17
Accepted: 2023-08-17
Published Online: 2023-09-13
Published in Print: 2024-01-26

© 2023 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2023-0518
Keywords for this article
digital immunoassay; neuronal injury; multiple sclerosis; validation

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. Rapid rule-in and rule-out protocols of acute myocardial infarction using hs-cTnI and hs-cTnT methods
  4. Reviews
  5. A guide to conducting systematic reviews of clinical laboratory tests
  6. Improving diagnosis and treatment of hypomagnesemia
  7. Opininon Paper
  8. Documenting and validating metrological traceability of serum alanine aminotransferase measurements: a priority for medical laboratory community for providing high quality service in hepatology
  9. EFLM Paper
  10. Assessing the status of European laboratories in evaluating biomarkers for chronic kidney diseases (CKD) and recommendations for improvement: insights from the 2022 EFLM Task Group on CKD survey
  11. General Clinical Chemistry and Laboratory Medicine
  12. Bland and Altman agreement method: to plot differences against means or differences against standard? An endless tale?
  13. Assessment of three equations to calculate plasma LDL cholesterol concentration in fasting and non-fasting hypertriglyceridemic patients
  14. Validation of metrological traceability of the new generation of Abbott Alinity alkaline phosphatase assay
  15. Quality assurance programs for vitamin A and E in serum: are we doing enough to assess laboratory performance?
  16. A supervised machine-learning approach for the efficient development of a multi method (LC-MS) for a large number of drugs and subsets thereof: focus on oral antitumor agents
  17. Performance evaluation of alternate ESR measurement method using BC-780 automated hematology analyzer: a comparison study with the Westergren reference method
  18. Total error in lymphocyte subpopulations by flow cytometry-based in state of the art using Spanish EQAS data
  19. Development and multi-center validation of a fully automated digital immunoassay for neurofilament light chain: toward a clinical blood test for neuronal injury
  20. CSF and plasma Aβ42/40 across Alzheimer’s disease continuum: comparison of two ultrasensitive Simoa® assays targeting distinct amyloid regions
  21. Improving regional medical laboratory center report quality through a report recall management system
  22. Cardiovascular Diseases
  23. Evaluation of the analytical and clinical performance of a new high-sensitivity cardiac troponin I assay: hs-cTnI (CLIA) assay
  24. Hemodialysis and biomarkers of myocardial infarction – a cohort study
  25. Letters to the Editor
  26. Chatbot GPT can be grossly inaccurate
  27. Blood collection in heparin vs. EDTA results in an inflammasome-independent increase in monocyte distribution width at 4 h
  28. Week-to-week within-subject and between-subject biological variation of copeptin
  29. The effect of unintended shortage in technical resources on the quality of endpoint clinical laboratory diagnosis
  30. Epigenetic signatures of cfDNA enable a topological assignment of tissue damage
  31. Six Sigma driven QC in antibody testing for infectious diseases
  32. Extensive analytical evaluation of the performances of the new DiaSys PCT assay and comparison with Elecsys B·R·A·H·M·S PCT test on Roche Cobas and B·R·A·H·M·S PCT-sensitive Kryptor
  33. Comment to: Extensive analytical evaluation of the performances of the new DiaSys PCT assay and comparison with Elecsys B·R·A·H·M·S PCT test on Roche Cobas and B·R·A·H·M·S PCT-sensitive Kryptor
  34. Evaluation of serum cortisol measurement by Ortho Vitro after stimulation by ACTH or insulin hypoglycaemia
  35. Diagnostic and prognostic value of quantitative detection of antimitochondrial antibodies subtype M2 using chemiluminescence immunoassay in primary biliary cholangitis
  36. Blood plasma biomarkers for Alzheimer’s disease: Aβ1–42/1–40 vs. AβX–42/X–40
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