A New Next-Generation Sequencing Strategy for the Simultaneous Analysis of Mutations and Chromosomal Rearrangements at Dna Level in Acute Myeloid Leukemia Patients

Publication date: Available online 9 October 2019Source: The Journal of Molecular DiagnosticsAuthor(s): M. Isabel Prieto-Conde, Luis A. Corchete, María García-Álvarez, Cristina Jiménez, Alejandro Medina, Ana Balanzategui, Montserrat Hernández-Ruano, Rebeca Maldonado, M. Eugenia Sarasquete, Miguel Alcoceba, Noemí Puig, Verónica González-Calle, Ramón García-Sanz, Norma C. Gutiérrez, Marcos González-Díaz, María Carmen ChillónAbstractAcute myeloid leukemias (AMLs) are currently genomically characterized by karyotype, fluorescence in situ hybridization (FISH), RQ-PCR, and DNA sequencing. Next-generation sequencing offers the promise of detecting all genomic lesions in a single run. However, technical limitations have hampered the detection of chromosomal rearrangements, so most studies are limited to somatic mutation assessment, or require the use of RNA-based strategies. To overcome these limitations, we designed a targeted-DNA capture next-generation sequencing approach associated with easy-to-perform public bioinformatic tools for one-step identification of translocations, inversions, and somatic mutations in AML. Thirty well-characterized newly diagnosed myeloid leukemia patients (27 AML and three chronic myeloid leukemia) were tested with the panel. Twenty-three of 24 known rearrangements, as well as one novel fusion gene that could not be detected by karyotype/FISH/RQ-PCR, were detected. This strategy also identified all chromosomal breakpoints as potential ta...
Source: The Journal of Molecular Diagnostics - Category: Pathology Source Type: research