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  • Biotechnical Methods Section BTS
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Biotechnical Methods Section

Simultaneous detection of NPM1 and FLT3-ITD mutations by capillary electrophoresis in acute myeloid leukemia

Leukemia volume 19, pages 1479–1482 (2005)Cite this article

A Corrigendum to this article was published on 25 April 2007

Abstract

Mutations in the Nucleophosmin (NPM1) gene have been recently described to occur in about one-third of acute myeloid leukemias (AML) and represent the most frequent genetic alteration currently known in this subset. These mutations generate an elongated NPM1 protein that localizes aberrantly in the cytoplasm. In analogy with Flt3 alterations, NPM1 mutations are mostly detectable in AML with normal karyotype and their recognition may be relevant to identify distinct response to treatment. Hence, in addition to conventional karyotyping and RT-PCR of fusion genes, combined analysis of both Flt3 and NPM1 mutations will be increasingly relevant in the genetic diagnosis work-up of AML. We developed a multiplex RT-PCR assay followed by capillary electrophoresis to simultaneously analyze NPM1 and Flt3 gene alterations (NFmPCR assay). The assay was validated in leukemic cell RNAs extracted from 38 AML patients, which had been previously characterized for Flt3 status by conventional RT-PCR. Direct sequencing of NPM1 RT-PCR products was carried out in 15 cases to verify results obtained by capillary electrophoresis. Both NPM1 sequencing and conventional RT-PCR Flt3 results showed 100% concordance with the results of the NFmPCR assay. We suggest that this assay may be introduced in routine analysis of genetic alterations in AML.

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Acknowledgements

This work was supported by MIUR Cofin 2003, and AIRC. At the time of the study, NIN was on leave of absence from the Department of Chemical Biochemistry (Hematology) Universidad Nacional de Rosario (Argentina) and EA was on leave of absence from the Division of Hematology of the University of Palermo (Italy). NIN is a member of Consejo Nacional de Investigaciones Cientificas y Técnicas (CONICET), Argentina.

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Authors and Affiliations

  1. Department of Biopathology, University of Tor Vergata, Rome, Italy

    N I Noguera, E Ammatuna, D Zangrilli, S Lavorgna, M Divona, F Buccisano, S Amadori & F Lo-Coco

  2. Institute of Hematology, University of Perugia, Perugia, Italy

    C Mecucci & B Falini

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  1. N I Noguera
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  2. E Ammatuna
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  4. S Lavorgna
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  9. B Falini
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  10. F Lo-Coco
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Correspondence to F Lo-Coco.

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Noguera, N., Ammatuna, E., Zangrilli, D. et al. Simultaneous detection of NPM1 and FLT3-ITD mutations by capillary electrophoresis in acute myeloid leukemia. Leukemia 19, 1479–1482 (2005). https://doi.org/10.1038/sj.leu.2403846

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