Denaturing high-performance liquid chromatography: a valid approach for identifying NPM1 mutations in acute myeloid leukemia.

The Journal of Molecular Diagnostics : JMD
Giovanni RotiGruppo Italiano Malattie Ematologiche dell' Adulto Working Party

Abstract

NPM1 gene mutations are the most frequent genetic lesion in the 60% of adult acute myeloid leukemias (AMLs) with normal karyotype and no evidence of typical fusion genes (BCR/ABL1, PML/RARA, AML1/ETO, CBFB/MYH11, DEK/CAN). Using direct sequencing we previously identified six different heterozygous mutants within exon 12 encoding the nucleophosmin C-terminus. Because of these mutations the shuttling protein nucleophosmin is aberrantly delocalized in the cytoplasm of leukemic cells (NPMc+). Here, we designed and tested a denaturing high-performance liquid chromatography (DHPLC) assay to detect NPM1 mutated variants. To assess specificity, sensitivity, reliability, and reproducibility, we analyzed DNA from 120 primary adult AMLs and compared DHPLC results with immunohistochemistry and sequencing. All electropherogram profiles in the 26 NPMc+ leukemias were different from the wild type, indicating 100% sensitivity. Sequencing categorized mutations A, B, and D, and all mutation A cases gave identical elution profiles. The other mutations showed typical chromatograms, with mutations B and D differing for one nucleotide. Elution profiles and sequencing also identified four new variants. Our results suggest that DHPLC detects NPM1mutat...Continue Reading

References

Jan 1, 1995·Cytogenetics and Cell Genetics·M HöglundF Mitelman
Jun 1, 2001·Human Mutation·W Xiao, P J Oefner
Dec 18, 2004·Current Opinion in Hematology·Guido MarcucciClara D Bloomfield
Apr 29, 2005·The New England Journal of Medicine·Masao NakagawaRitsuro Suzuki

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Citations

Sep 3, 2011·International Journal of Laboratory Hematology·A J Bench
Jan 9, 2008·Leukemia Research·Meilani SyampurnawatiYoshitake Hayashi
Aug 22, 2012·Acta Haematologica·Milos KuzmanovicMilica Colovic
Sep 19, 2009·Expert Review of Anticancer Therapy·Natalia Meani, Myriam Alcalay
Dec 17, 2015·Asia-Pacific Journal of Clinical Oncology·Pulkit RastogiSubhash Varma
Feb 6, 2018·Indian Journal of Hematology & Blood Transfusion : an Official Journal of Indian Society of Hematology and Blood Transfusion·Jina BhattacharyyaDushyant Kumar
Dec 3, 2013·Asian Pacific Journal of Cancer Prevention : APJCP·Sadaf ShahabNuzhat Ahmed
Dec 5, 2009·The Journal of Molecular Diagnostics : JMD·Margaret L GulleyYuri Fedoriw

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