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Bio-Technical Methods Section

Validation of BIOMED-2 multiplex PCR tubes for detection of TCRB gene rearrangements in T-cell malignancies

Leukemia volume 18, pages 1531–1538 (2004)Cite this article

Abstract

The BIOMED-2 Concerted Action BMH4-CT98-3936 on ‘Polymerase chain reaction (PCR)-based clonality studies for early diagnosis of lymphoproliferative disorders’ developed standardized PCR protocols for detection of immunoglobulin (Ig) and T-cell receptor (TCR) rearrangements, including TCR beta (TCRB). As no comparable TCRB PCR method pre-existed and only a limited number of samples was tested within the BIOMED-2 study, we initiated this study for further validation of the newly developed TCRB PCR approach by comparing PCR data with previously generated Southern blot (SB) data in a series of 66 immature (ALL) and 36 mature T-cell malignancies. In 91% of cases, concordant PCR and SB results were found. Discrepancies consisted of either failure to detect SB-detected TCRB rearrangements by PCR (6.5%) or detection of an additional non-SB defined rearrangement (2.5%). In 99% of cases (99/100), at least one clonal TCRB rearrangement was detected by PCR in the SB-positive cases. A predominance of complete Vβ-Jβ rearrangements was seen in TCRαβ+ T-cell malignancies and CD3-negative T-ALL (100 and 90%, respectively), whereas in TCRγδ+ T-ALL, more incomplete Dβ-Jβ TCRB rearrangements were detected (73%). Our results underline the reliability of this new TCRB PCR method and its strategic applicability in clonality diagnostics of lymphoproliferative disorders and MRD studies.

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Acknowledgements

We gratefully acknowledge Petra Chall, Frauke Hemken, Ellen van Gastel-Mol for their technical assistance and Mr Tar van Os for help in preparation of the figure. This work was supported by the BIOMED-2 Concerted Action BMH4-CT98-3936 ‘PCR-based clonality studies for early diagnosis of lymphoproliferative disorders’ of the European Commission.

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Authors and Affiliations

  1. Medical Clinic II, University of Kiel, Germany

    J Droese, M Brüggemann, P Neumann & M Kneba

  2. Department of Immunology, Erasmus MC, University Medical Center Rotterdam, The Netherlands

    A W Langerak, I L M Wolvers-Tettero & J J M van Dongen

  3. Department of Pathology, University Medical Center St Radboud Nijmegen, The Netherlands

    P J T A Groenen & M C van Altena

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  1. J Droese
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  2. A W Langerak
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  6. I L M Wolvers-Tettero
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Correspondence to J J M van Dongen.

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Droese, J., Langerak, A., Groenen, P. et al. Validation of BIOMED-2 multiplex PCR tubes for detection of TCRB gene rearrangements in T-cell malignancies. Leukemia 18, 1531–1538 (2004). https://doi.org/10.1038/sj.leu.2403428

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