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  • Biotechnical Methods Section BTS
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Biotechnical Methods Section (BTS)

Paired multiplex reverse-transcriptase polymerase chain reaction (PMRT-PCR) analysis as a rapid and accurate diagnostic tool for the detection of MLL fusion genes in hematologic malignancies

Leukemia volume 15, pages 1293–1300 (2001)Cite this article

Abstract

The MLL gene in chromosome band 11q23 is frequently rearranged in acute lymphoblastic and acute myeloid leukemias. To date, more than 50 different chromosomal regions are known to participate in translocations involving 11q23, many of which affect MLL. The pathogenetically important outcome of these rearrangements is most likely the creation of a fusion gene consisting of the 5′ part of the MLL gene and the 3′ end of the partner gene. Although abnormalities of the MLL gene as such are generally associated with poor survival, recent data suggest that the prognostic impact varies among the different fusion genes generated. Hence, detection of the specific chimeric gene produced is important for proper prognostication and clinical decision making. We have developed a paired multiplex reverse-transcriptase polymerase chain reaction analysis to facilitate a rapid and accurate detection of the most frequent MLL fusion genes in adult and childhood acute leukemias. To increase the specificity, two sets of primers were designed for each fusion gene, and these paired primer sets were run in parallel in two separate multiplex one-step PCR reactions. Using the described protocol, we were able to amplify successfully, in one single assay, the six clinically relevant fusion genes generated by the t(4;11)(q21;q23) [MLL/AF4], t(6;11)(q27;q23) [MLL/AF6], t(9;11)(p21–22;q23) [MLL/AF9], t(10;11)(p11–13;q23) [MLL/AF10], t(11;19)(q23;p13.1) [MLL/ELL], and t(11;19)(q23; p13.3) [MLL/ENL] in cell lines, as well as in patient material.

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Acknowledgements

This study was supported by grants from the Swedish Cancer Society, the Swedish Child Cancer Fund, and the Lund University Medical Faculty.

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  1. Department of Clinical Genetics, Lund University Hospital, Sweden

    A Andersson, M Höglund, B Johansson, C Lassen, F Mitelman & T Fioretos

  2. Department of Internal Medicine, Division of Hematology, Lund University Hospital, Sweden

    R Billström & PG Nilsson

  3. Department of Pediatrics, Lund University Hospital, Sweden

    S Garwicz

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Andersson, A., Höglund, M., Johansson, B. et al. Paired multiplex reverse-transcriptase polymerase chain reaction (PMRT-PCR) analysis as a rapid and accurate diagnostic tool for the detection of MLL fusion genes in hematologic malignancies. Leukemia 15, 1293–1300 (2001). https://doi.org/10.1038/sj.leu.2402189

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