A new MLPA-based method for the detection of acute myeloid leukemia-specific NPM1 mutations and simultaneous analysis of gene copy number alterations

Publication date: Available online 4 February 2016 Source:Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis Author(s): Malgorzata Marcinkowska-Swojak, Luiza Handschuh, Pawel Wojciechowski, Michal Goralski, Kamil Tomaszewski, Maciej Kazmierczak, Krzysztof Lewandowski, Mieczyslaw Komarnicki, Jacek Blazewicz, Marek Figlerowicz, Piotr Kozlowski The NPM1 gene encodes nucleophosmin, a protein involved in multiple cell functions and carcinogenesis. Mutation of the NPM1 gene, causing delocalization of the protein, is the most frequent genetic lesion in acute myeloid leukemia (AML); it is considered a founder event in AML pathogenesis and serves as a favorable prognostic marker. Moreover, in solid tumors and some leukemia cell lines, overexpression of the NPM1 gene is commonly observed. Therefore, the purpose of this study was to develop a new method for the detection of NPM1 mutations and the simultaneous analysis of copy number alterations (CNAs), which may underlie NPM1 gene expression deregulation. To address both of the issues, we applied a strategy based on multiplex ligation-dependent probe amplification (MLPA). A designed NPM1mut+ assay enables the detection of three of the most frequent NPM1 mutations: A, B and D. The accuracy of the assay was tested using a group of 83 samples from Polish patients with AML and other blood-proliferative disorders. To verify the results, we employed traditional Sanger sequencing and next-generation...
Source: Mutation Research Fundamental and Molecular Mechanisms of Mutagenesis - Category: Cytology Source Type: research