Detecting minimal residual disease in patients with chronic lymphocytic leukemia using 8-color flow cytometry protocol in routine hematological practice

Authors

  • O. Stehlíková,

    1. Department of Internal Medicine – Hematology and Oncology, Brno and Faculty of Medicine, University Hospital, Masaryk University, Brno, Czech Republic
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    • These authors contributed equally to this work.
  • J. Chovancová,

    1. Department of Internal Medicine – Hematology and Oncology, Brno and Faculty of Medicine, University Hospital, Masaryk University, Brno, Czech Republic
    2. Central European Institute of Technology, Masaryk University, Brno, Czech Republic
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    • These authors contributed equally to this work.
  • B. Tichý,

    1. Department of Internal Medicine – Hematology and Oncology, Brno and Faculty of Medicine, University Hospital, Masaryk University, Brno, Czech Republic
    2. Central European Institute of Technology, Masaryk University, Brno, Czech Republic
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  • M. Krejčí,

    1. Department of Internal Medicine – Hematology and Oncology, Brno and Faculty of Medicine, University Hospital, Masaryk University, Brno, Czech Republic
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  • Y. Brychtová,

    1. Department of Internal Medicine – Hematology and Oncology, Brno and Faculty of Medicine, University Hospital, Masaryk University, Brno, Czech Republic
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  • A. Panovská,

    1. Department of Internal Medicine – Hematology and Oncology, Brno and Faculty of Medicine, University Hospital, Masaryk University, Brno, Czech Republic
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  • H. Francová Skuhrová,

    1. Department of Internal Medicine – Hematology and Oncology, Brno and Faculty of Medicine, University Hospital, Masaryk University, Brno, Czech Republic
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  • K. Burčková,

    1. Department of Internal Medicine – Hematology and Oncology, Brno and Faculty of Medicine, University Hospital, Masaryk University, Brno, Czech Republic
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  • M. Borský,

    1. Department of Internal Medicine – Hematology and Oncology, Brno and Faculty of Medicine, University Hospital, Masaryk University, Brno, Czech Republic
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  • T. Loja,

    1. Department of Internal Medicine – Hematology and Oncology, Brno and Faculty of Medicine, University Hospital, Masaryk University, Brno, Czech Republic
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  • J. Mayer,

    1. Department of Internal Medicine – Hematology and Oncology, Brno and Faculty of Medicine, University Hospital, Masaryk University, Brno, Czech Republic
    2. Central European Institute of Technology, Masaryk University, Brno, Czech Republic
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  • Š. Pospíšilová,

    1. Department of Internal Medicine – Hematology and Oncology, Brno and Faculty of Medicine, University Hospital, Masaryk University, Brno, Czech Republic
    2. Central European Institute of Technology, Masaryk University, Brno, Czech Republic
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  • M. Doubek

    Corresponding author
    1. Department of Internal Medicine – Hematology and Oncology, Brno and Faculty of Medicine, University Hospital, Masaryk University, Brno, Czech Republic
    2. Central European Institute of Technology, Masaryk University, Brno, Czech Republic
    • Correspondence:

      Michael Doubek, University Hospital and Masaryk University, Jihlavská 20, 62500 Brno, Czech Republic. Tel.: +42 0 532 233 642; Fax: + 42 0 532 233 603; E-mail: mdoubek@fnbrno.cz

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Summary

Introduction

Minimal residual disease (MRD) detection has become increasingly important for the assessment of therapy response in chronic lymphocytic leukemia (CLL). However, current MRD analysis methods, both molecular genetic and flow cytometric, are time-consuming and require experienced laboratory staff.

Methods

To reduce the demands of flow cytometric MRD detection in CLL, we have introduced a novel flow cytometric 8-color protocol. The MRD analysis results using this protocol were then compared with the commonly employed 4-color protocol and the molecular genetic (real-time quantitative allele-specific oligonucleotide IGH polymerase chain reaction; RQ-ASO IGH PCR) approach.

Results

Forty-two CLL patient samples were repeatedly analyzed after allogeneic stem cell transplantation (n = 20) or after fludarabine-based therapy (n = 22), and 100% concordance was found using both flow cytometric protocols. Furthermore, there was a strong correlation (r = 0.94) between flow cytometric and RQ-ASO IGH PCR results in MRD detection.

Conclusion

Flow cytometry is less time-consuming, less financially demanding, and moreover, MRD assessment using our novel 8-color protocol is less complicated than the 4-color approach and molecular methods.

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