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Rapid identification of frequent MLL rearrangements in hematolgic malignancies by multiplex RT-PCR in a single assay

Leukemia volume 16, page 1574 (2002)Cite this article

Andersson et al1 reported the development of a paired multiplex reverse-transcriptase polymerase chain reaction (PMRT-PCR) for the detection of MLL fusion genes in hematologic malignancies. They described this method as allowing rapid and accurate detection of the clinically relevant fusion genes MLL/AF4, MLL/AF6, MLL/AF9, MLL/AF10, MLL/ELL and MLL/ENL in one single assay. In their multiplex assay they applied two separate primer pairs, for each fusion gene, that were used in two parallel one-step multiplex PCR reactions with ‘out-mix’ primers and ‘in-mix’ primers. Thus each sample was tested twice for the presence of a specific MLL fusion gene and the designed out-mixes and in-mixes that produced PCR fragments increased the specificity of the assay. However, a split-out reaction for each case was necessary in a second step to identify the specific fusion genes with corresponding primers.

We had developed in our laboratory a corresponding strategy using PMRT-PCR for the detection of MLL fusion genes but we were able to identify the partner gene only in one single assay. In our protocol we analyzed the frequent fusion genes involved in acute myeloid leukemia (LAM) as AF6, AF9 and ELL and in acute lymphoblastic leukemia (LAL) as AF4 and ENL.

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References

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Acknowledgements

This work was supported by the regional Hospital Center of Montpellier. I thank Dr Jean Marc Rey for sequencing and Jean Pierre Bonifas for technical assistance.

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  1. Génétique Moléculaire et Chromosomique Hôpital A de Villeneuve Montpellier, France

    M Dupont

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  1. M Dupont
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Dupont, M. Rapid identification of frequent MLL rearrangements in hematolgic malignancies by multiplex RT-PCR in a single assay. Leukemia 16, 1574 (2002). https://doi.org/10.1038/sj.leu.2402560

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