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Chronic Lymphocytic Leukemia

Standardized MRD flow and ASO IGH RQ-PCR for MRD quantification in CLL patients after rituximab-containing immunochemotherapy: a comparative analysis

Leukemia volume 23, pages 2007–2017 (2009)Cite this article

Abstract

Rituximab-containing regimens are becoming a therapeutic standard in chronic lymphocytic leukemia (CLL), so that a validation of flow cytometric minimal residual disease (MRD) quantification (MRD flow) in the presence of this antibody is necessary. We therefore compared results obtained by real-time quantitative (RQ)-PCR to MRD flow in 530 samples from 69 patients randomized to receive chemotherapy or chemotherapy plus rituximab. Quantitative MRD levels assessed by both techniques were closely correlated irrespective of therapy (r=0.95). The sensitivity and specificity of MRD flow was not influenced by the presence of rituximab. With 58.9% positive and 26.4% negative samples by both techniques, 85.3% of assessments (452/530) were qualitatively concordant between MRD flow and RQ-PCR. Discordant samples were typically negative by MRD flow and simultaneously positive close to the detection limit of the PCR assays, indicating a higher sensitivity of PCR for very low MRD levels. However, 93.8% of all samples were concordantly classified by both methods using a threshold of 10−4 to determine MRD positivity. MRD flow and PCR are equally effective for MRD quantification in rituximab-treated CLL patients within a sensitivity range of up to 10−4, whereas PCR is more sensitive for detecting MRD below that level.

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Acknowledgements

We are grateful to the German CLL Study Group (GCLLSG) and the contributing centers taking care of patients being enrolled in this study. The contribution of the GCLLSG study center (Anne Westermann, Anna-Maria Fink) is gratefully acknowledged. We thank Elke Harbst, Vanessa Krull, Jamileh Hanani, Maike Starken, Lada Henseleit, Sebastian Buske, Martina Klingenberg and Karin Olsen for excellent technical assistance. We also thank F Hoffmann—La Roche Ltd, Basel, for research funding provided.

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Authors and Affiliations

  1. Second Department of Medicine, University of Schleswig-Holstein, Kiel, Schleswig-Holstein, Germany

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

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

    S Stilgenbauer & H Döhner

  3. Institute of Medical Statistics and Epidemiology, Technical University, Munich, Germany

    R Busch

  4. Department I of Internal Medicine, and Center of Integrated Oncology Cologne Bonn, University Hospital, Cologne, Germany

    K Fischer, G Fingerle-Rowson & M Hallek

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Böttcher, S., Stilgenbauer, S., Busch, R. et al. Standardized MRD flow and ASO IGH RQ-PCR for MRD quantification in CLL patients after rituximab-containing immunochemotherapy: a comparative analysis. Leukemia 23, 2007–2017 (2009). https://doi.org/10.1038/leu.2009.140

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