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Chronic Lymphocytic Leukemias

A new automated flow cytometry data analysis approach for the diagnostic screening of neoplastic B-cell disorders in peripheral blood samples with absolute lymphocytosis

Leukemia volume 20, pages 1221–1230 (2006)Cite this article

Abstract

Currently, multiparameter flow cytometry immunophenotyping is the selected method for the differential diagnostic screening between reactive lymphocytosis and neoplastic B-cell chronic lymphoproliferative disorders (B-CLPD). Despite this, current multiparameter flow cytometry data analysis approaches still remain subjective due to the need of experienced personnel for both data analysis and interpretation of the results. In this study, we describe and validate a new automated method based on vector quantization algorithms to analyze multiparameter flow cytometry immunophenotyping data in a series of 307 peripheral blood (PB) samples. Our results show that the automated method of analysis proposed compares well with currently used manual approach and significantly improves semiautomated approaches and, that by using it, a highly efficient discrimination with 100% specificity and 100% sensitivity can be made between normal/reactive PB samples and cases with B-CLPD based on the total B-cell number and/or the sIgκ+/sIgλ+ B-cell ratio. In addition, the method proved to be able to detect the presence of pathologic neoplastic B-cells even when these are present at low frequencies (<5% of all lymphocytes in the sample) and in poor-quality samples enriched in ‘noise’ events.

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Acknowledgements

This work has been partially supported by grants from the Ministerio de Educación y Ciencia SAF 2002-03096, the Spanish Network of Cancer Research Centers (Ref C03/10) (FIS, Ministerio de Sanidad y Consumo) and Programa Hispano-Brasileño de Cooperación Universitaria Ref. PHB 2004-0800-PC (Ministerio de Educación y Ciencia), Madrid, Spain, and CAPES/Ministerio da Educação, Brasília, Brazil. MDT is supported by a grant from Ministerio de Ciencia y Tecnología (Programa Ramón y Cajal), Madrid, Spain. CEP was partially supported by a grant from CNPq- Brazilian National Research Council, Brasília. Brazil.

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Author notes

  1. E S Costa and M E Arroyo: These authors contributed equally to this work.

Authors and Affiliations

  1. Instituto de Pediatria e Puericultura Martagão Gesteira and Departamento de Clínica Médica, URFJ/Federal University of Rio de Janeiro, Rio de Janeiro, Brazil

    E S Costa

  2. Department of Medicine and Cancer Research Center, Cytometry Service, University of Salamanca, Salamanca, Spain

    M E Arroyo, J Almeida & A Orfao

  3. Faculty of Medicine and COPPE – Engineering Graduate Program, URFJ/Federal University of Rio de Janeiro, Rio de Janeiro, Brazil

    C E Pedreira

  4. Hematology Service, University Hospital of Salamanca, Salamanca, Spain

    M A García-Marcos

  5. Research Unity, University Hospital of Salamanca, Salamanca, Spain

    M D Tabernero

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  1. E S Costa
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  2. M E Arroyo
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  3. C E Pedreira
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  4. M A García-Marcos
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  5. M D Tabernero
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  6. J Almeida
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  7. A Orfao
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Correspondence to A Orfao.

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Costa, E., Arroyo, M., Pedreira, C. et al. A new automated flow cytometry data analysis approach for the diagnostic screening of neoplastic B-cell disorders in peripheral blood samples with absolute lymphocytosis. Leukemia 20, 1221–1230 (2006). https://doi.org/10.1038/sj.leu.2404241

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