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  • Biotechnical Methods Section BTS
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Biotechnical Methods Section (BTS)

Detection of translocations affecting the BCL6 locus in B cell non-Hodgkin's lymphoma by interphase fluorescence in situ hybridization

Leukemia volume 15, pages 1475–1484 (2001)Cite this article

Abstract

Structural alterations in 3q27 affecting the BCL6 locus are among the most frequent changes in B-NHL. The aim of the present study was to establish an interphase-FISH assay for the detection of all diverse BCL6 translocations in B-NHL. Two different approaches were tested, one using a PAC-clone spanning the major breakpoint region (MBR) of BCL6(span-assay), and another using two BAC clones flanking the MBR (flank-assay). Interphase FISH with the span-assay detected the various BCL6 translocations in seven B-NHL cell lines. The dual-color flank-assay was evaluated in two laboratories independently: in normal controls, the cutoff level for false-positive signals was 2.6%, whereas the cutoff level for false-negatives in the seven cell lines was 7.5%. To test the feasibility of the FISH strategies, 30 samples from patients with B-NHL with cytogenetic abnormalities of 3q27 were evaluated with both assays. In 21 cases, the span-assay indicated a BCL6 rearrangement. In 18 of the 21 cases, the dual-color flank-assay confirmed the translocation including 12 different partner chromosomal loci. The three false-positive cases detected with the span-assay showed trisomy of chromosome 3 by cytogenetic analyses, and they were correctly classified as non-rearranged with the flank-assay. In summary, our FISH strategy using two differently labeled flanking BCL6 BAC probes provides a robust, sensitive, and reproducible method for the detection of common and uncommon abnormalities of BCL6 gene in interphase nuclei. The routine application of this assay to patients with B-NHL will allow the assessment of the diagnostic and prognostic significance of BCL6 rearrangements.

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Acknowledgements

This work was supported by grants from the Fondo de Investigación Sanitaria FIS-98/0491 and BECE-99/6023, by the Generalitat Valenciana, Conselleria de Educacion y Ciencia POST-99/12–61, by the Deutsche Krebshilfe Grants 10–1556-Schl4 and 10–1641-De1, and by the IZKF Kiel. We thank Gianluca Gaidano (Torino, Italy), for the OCI-Ly8 cell line; Hitoshi Ohno (Kyoto, Japan), for the YM cell line; Wen-Lin Kuo and Joe Grey (University of California, San Francisco) for PACs RMC03P056 and RMC03P061; Mariano Rocci (Bari, Italy), for PACs 1019H10 and 1098H17; and M Ordoñez, E Cervello, R Marques, M Botia, A Schneider, F Kahlert, L Askari, C Becher, D Schuster and R Zühlke-Jenisch for excellent technical assistance.

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Authors and Affiliations

  1. Department of Hematology and Oncology, Hospital Clinico, University of Valencia, Spain

    D Sanchez-Izquierdo, I Marugan, I Benet, J Garcia-Conde & JA Martinez-Climent

  2. Institute of Human Genetics, University Hospital Kiel, Germany

    R Siebert, L Harder, S Gesk & B Schlegelberger

  3. Section of Hematology and Oncology, University of Chicago, USA

    A Gozzetti & MM Le Beau

  4. Academic Dept of Haematology and Cytogenetics, Institute of Cancer Research, Sutton, UK

    HP Price, T Sonoki & MJS Dyer

  5. Department of Hematology, Hospital Clínico, University of Salamanca, Spain

    JM Hernandez-Rivas

  6. Department of Hematology, Hospital del Mar, Barcelona, Spain

    F Solé

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  1. D Sanchez-Izquierdo
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Sanchez-Izquierdo, D., Siebert, R., Harder, L. et al. Detection of translocations affecting the BCL6 locus in B cell non-Hodgkin's lymphoma by interphase fluorescence in situ hybridization. Leukemia 15, 1475–1484 (2001). https://doi.org/10.1038/sj.leu.2402207

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