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  • Biotechnical Methods Section BTS
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Bio-Technical Methods Section

A diagnostic biochip for the comprehensive analysis of MLL translocations in acute leukemia

Leukemia volume 18, pages 1522–1530 (2004)Cite this article

Abstract

Reciprocal rearrangements of the MLL gene are among the most common chromosomal abnormalities in both Acute Lymphoblastic and Myeloid Leukemia. The MLL gene, located on the 11q23 chromosomal band, is involved in more than 40 recurrent translocations. In the present study, we describe the development and validation of a biochip-based assay designed to provide a comprehensive molecular analysis of MLL rearrangements when used in a standard clinical pathology laboratory. A retrospective blind study was run with cell lines (n=5), and MLL positive and negative patient samples (n=31), to evaluate assay performance. The limits of detection determined on cell line data were 10−1, and the precision studies yielded 100% repeatability and 98% reproducibility. The study shows that the device can detect frequent (AF4, AF6, AF10, ELL or ENL) as well as rare partner genes (AF17, MSF). The identified fusion transcripts can then be used as molecular phenotypic markers of disease for the precise evaluation of minimal residual disease by RQ-PCR. This biochip-based molecular diagnostic tool allows, in a single experiment, rapid and accurate identification of MLL gene rearrangements among 32 different fusion gene (FG) partners, precise breakpoint positioning and comprehensive screening of all currently characterized MLL FGs.

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Acknowledgements

We thank Dr C Bilhou Nabera (CHU, Bordeaux, France) and Dr N Dastugue (Hôpital Purpan, Toulouse, France) for providing samples, Dr D Birnbaum (IPC, Marsèille, France) for critical review of the manuscript, and Dr P Ravassard (LGN, Paris, France) for valuable discussion. We also gratefully acknowledge Ipsogen's Bioinformatics and Production Teams for their contributions. This work was partially supported by grants from PHRC97 (JG), the Agence Nationale de la Valorisation de la Recherche (ANVAR) and the Directions Régionales de l'Industrie de la Recherche et de l'Environnement (DRIRE).

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Authors and Affiliations

  1. IPSOGEN SAS, Case 923, 163, Av. de Luminy, Marseille, Cedex 9, France

    N Maroc, A Morel, A L de La Chapelle, A Koki, V Fert & F Hermitte

  2. ERT MEIDIA, IFR Jean Roche, Marseille, Cedex 20, France

    E Beillard & J Gabert

  3. INSERM U462, Centre Hayem, Hôpital Saint Louis, Paris, Cedex 10, France

    X Fund & J-M Cayuela

  4. Laboratoire de cytogénétique, Institut Paoli-Calmettes, 232, bd Ste Marguerite, Marseille, France

    M-J Mozziconacci

  5. LGMC Hôpital Arnaud de Villeneuve, Montpellier, France

    M Dupont

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  1. N Maroc
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Correspondence to N Maroc.

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Maroc, N., Morel, A., Beillard, E. et al. A diagnostic biochip for the comprehensive analysis of MLL translocations in acute leukemia. Leukemia 18, 1522–1530 (2004). https://doi.org/10.1038/sj.leu.2403439

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