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Minimal Residual Disease in CLL

Comparative analysis of minimal residual disease detection using four-color flow cytometry, consensus IgH-PCR, and quantitative IgH PCR in CLL after allogeneic and autologous stem cell transplantation

Leukemia volume 18, pages 1637–1645 (2004)Cite this article

Abstract

The clinically most suitable method for minimal residual disease (MRD) detection in chronic lymphocytic leukemia is still controversial. We prospectively compared MRD assessment in 158 blood samples of 74 patients with CLL after stem cell transplantation (SCT) using four-color flow cytometry (MRD flow) in parallel with consensus IgH-PCR and ASO IgH real-time PCR (ASO IgH RQ-PCR). In 25 out of 106 samples (23.6%) with a polyclonal consensus IgH-PCR pattern, MRD flow still detected CLL cells, proving higher sensitivity of flow cytometry over PCR-genescanning with consensus IgH-primers. Of 92 samples, 14 (15.2%) analyzed in parallel by MRD flow and by ASO IgH RQ-PCR were negative by our flow cytometric assay but positive by PCR, thus demonstrating superior sensitivity of RQ-PCR with ASO primers. Quantitative MRD levels measured by both methods correlated well (r=0.93). MRD detection by flow and ASO IgH RQ-PCR were equally suitable to monitor MRD kinetics after allogeneic SCT, but the PCR method detected impending relapses after autologous SCT earlier. An analysis of factors that influence sensitivity and specificity of flow cytometry for MRD detection allowed to devise further improvements of this technique.

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Acknowledgements

We are grateful to the German CLL Study Group (DCLLSG; chairman Professor Michael Hallek) and the contributing centers taking care of patients being enrolled in this study.

We also thank Elke Harbst, Karin Brune, Christa Waubke, Daniela Krüss, Alexandra Lange, and Christian Bretscher for excellent technical assistance. This study was supported by ‘Deutsche José Carreras Leukämie-Stiftung e.V.’ (LR02/18, R16) and by ‘Fresenius Stiftung’.

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Authors and Affiliations

  1. Second Department of Medicine, University of Kiel, Germany

    S Böttcher, M Ritgen, C Pott, M Brüggemann, T Raff & M Kneba

  2. Third Department of Internal Medicine, University of Ulm, Germany

    S Stilgenbauer & H Döhner

  3. Department of Hematology, AK St Georg, Hamburg, Germany

    P Dreger

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  1. S Böttcher
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Correspondence to S Böttcher.

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Böttcher, S., Ritgen, M., Pott, C. et al. Comparative analysis of minimal residual disease detection using four-color flow cytometry, consensus IgH-PCR, and quantitative IgH PCR in CLL after allogeneic and autologous stem cell transplantation. Leukemia 18, 1637–1645 (2004). https://doi.org/10.1038/sj.leu.2403478

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