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Evaluation of analytical performance of AQUIOS CL flow cytometer and method comparison with bead-based flow cytometry methods

  • Andrada S. Chiron ORCID logo , Lucy Locher , Aurélie Sarthou , Aude Gleizes , Roman Krzysiek , Pascale Chretien and Salima Hacein-Bey-Abina EMAIL logo
Published/Copyright: April 8, 2024
Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 62 Issue 10

Abstract

Objectives

Given that method validation is mandatory for compliance with the International Organization for Standardization (ISO) 15,189 standard requirements, we evaluated the analytical performance of the AQUIOS CL system (Beckman Coulter) and compared it with two bead-based flow cytometry (FCM) protocols (BD FACSCAntoTM-II and Beckman Coulter DxFLEX). There are no comparative literature data on standardized protocols for counting lymphocyte subsets on the new-generation cytometer DxFLEX.

Methods

We evaluated the AQUIOS CL’s performance with regard to accuracy, linearity and stability by using dedicated control cell samples and patient samples. We also compared the lymphocyte counts measured on the AQUIOS CL (n=69 samples) with those measured on the BD FACSCAntoTM-II and DxFLEX FCM systems. For 61 samples, FCM results were compared with those measured on the XN-3000 Sysmex hematology analyzer.

Results

AQUIOS CL showed acceptable performance – even outside the manufacturer’s quantification ranges- and strong correlations with bead-based FCM methods. The FCM techniques and the XN-3000 gave similar absolute lymphocyte counts, although values in samples with intense lymphocytosis (B cell lymphoma/leukemia) were underestimated.

Conclusions

The AQUIOS CL flow cytometer is a time-saving, single-platform system with good performance, especially when the manufacturer’s instructions for use are followed. However, AQUIOS CL’s possible limitations and pitfalls impose validation of a bead-based FCM method for immunophenotyping verification or as a back-up system. Although the DxFLEX flow cytometer is more time-consuming to use, it can provide standardized lymphocyte subset counts in case of aberrant results on AQUIOS CL or in the event of equipment failure.

Keywords: lymphocyte subsets count; flow cytometry; AQUIOS CL; method validation
Corresponding author: Salima Hacein-Bey-Abina, Clinical Immunology Laboratory, Groupe Hospitalier Universitaire Paris Saclay, Hôpital Bicêtre, Assistance Publique-Hôpitaux de Paris, 78 Rue du Général Leclerc, Le Kremlin-Bicêtre, 94270, 94270, Le Kremlin-Bicêtre, France; and UTCBS, Unité des technologies Chimiques et Biologiques pour la Santé, Université Paris Cité, CNRS 8258, INSERM U1267, 4 Avenue de l’Observatoire, 75006, Paris, France, E-mail:

Acknowledgments

The authors thank all the staff in the Immunology Laboratory, Kremlin Bicêtre Hospital for their work during the study. We also thank staff in the hospital’s Hematology Laboratory for access to the Sysmex XN-3000 analyzer. Furthermore, we thank Beckman Coulter (and Annabelle Chauveau in particular) for technical assistance with the AQUIOS CL, DxFLEX and Kaluza C. Lastly, we thank Dr. David Fraser (Biotech Communication SARL, Ploudalmézeau, France) for copy-editing assistance.

  1. Research ethics: Not applicable.

  2. Informed consent: In accordance with our local Ethics Committee’s rules, there was no need for signed informed consent.

  3. Author contributions: SHBA, PC, AC: study design, SHBA, AC, PC: drafted the manuscript. AC, LL, AS: patient recruitment. LL, AS, AC: data acquisition and Kaluza analysis. AC, PC, SHBA, RK: statistical analysis, RK, SHBA, AG: revision of the manuscript’s structure, style and content. PC, AG: commented on and edited the manuscript, which was read and approved by all authors. All the authors have accepted responsibility for the entire content of this submitted manuscript and approved its submission.

  4. Competing interests: The authors state no conflict of interest.

  5. Research funding: None declared.

  6. Data availability: The raw data can be obtained on request from the corresponding author.

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Supplementary Material

This article contains supplementary material (https://doi.org/10.1515/cclm-2023-1498).

Received: 2023-12-27
Accepted: 2024-03-06
Published Online: 2024-04-08
Published in Print: 2024-09-25

© 2024 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2023-1498
Keywords for this article
lymphocyte subsets count; flow cytometry; AQUIOS CL; method validation

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. Six years of progress – highlights from the IFCC Emerging Technologies Division
  4. IFCC Papers
  5. Skin in the game: a review of single-cell and spatial transcriptomics in dermatological research
  6. Bilirubin measurements in neonates: uniform neonatal treatment can only be achieved by improved standardization
  7. Validation and verification framework and data integration of biosensors and in vitro diagnostic devices: a position statement of the IFCC Committee on Mobile Health and Bioengineering in Laboratory Medicine (C-MBHLM) and the IFCC Scientific Division
  8. Linearity assessment: deviation from linearity and residual of linear regression approaches
  9. HTA model for laboratory medicine technologies: overview of approaches adopted in some international agencies
  10. Considerations for applying emerging technologies in paediatric laboratory medicine
  11. A global perspective on the status of clinical metabolomics in laboratory medicine – a survey by the IFCC metabolomics working group
  12. The LEAP checklist for laboratory evaluation and analytical performance characteristics reporting of clinical measurement procedures
  13. General Clinical Chemistry and Laboratory Medicine
  14. Assessing post-analytical phase harmonization in European laboratories: a survey promoted by the EFLM Working Group on Harmonization
  15. Potential medical impact of unrecognized in vitro hypokalemia due to hemolysis: a case series
  16. Quantification of circulating alpha-1-antitrypsin polymers associated with different SERPINA1 genotypes
  17. Targeted ultra performance liquid chromatography tandem mass spectrometry procedures for the diagnosis of inborn errors of metabolism: validation through ERNDIM external quality assessment schemes
  18. Improving protocols for α-synuclein seed amplification assays: analysis of preanalytical and analytical variables and identification of candidate parameters for seed quantification
  19. Evaluation of analytical performance of AQUIOS CL flow cytometer and method comparison with bead-based flow cytometry methods
  20. IgG and kappa free light chain CSF/serum indices: evaluating intrathecal immunoglobulin production in HIV infection in comparison with multiple sclerosis
  21. Reference Values and Biological Variations
  22. Reference intervals of circulating secretoneurin concentrations determined in a large cohort of community dwellers: the HUNT study
  23. Sharing reference intervals and monitoring patients across laboratories – findings from a likely commutable external quality assurance program
  24. Verification of bile acid determination method and establishing reference intervals for biochemical and haematological parameters in third-trimester pregnant women
  25. Confounding factors of the expression of mTBI biomarkers, S100B, GFAP and UCH-L1 in an aging population
  26. Cancer Diagnostics
  27. Exploring evolutionary trajectories in ovarian cancer patients by longitudinal analysis of ctDNA
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  29. Evaluation of effects from hemoglobin variants on HbA1c measurements by different methods
  30. Letters to the Editor
  31. Are there any reasons to use three levels of quality control materials instead of two and if so, what are the arguments?
  32. Issues for standardization of neonatal bilirubinemia: a case of delayed phototherapy initiation
  33. The routine coagulation assays plasma stability – in the wake of the new European Federation of Clinical Chemistry and Laboratory Medicine (EFLM) biological variability database
  34. Improving HCV diagnosis following a false-negative anti-HCV result
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