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Type: Journal article
Title: A method for next-generation sequencing of paired diagnostic and remission Samples to detect mitochondrial DNA mutations associated with leukemia
Author: Pagani, I.
Kok, C.
Saunders, V.
Van der Hoek, M.
Heatley, S.
Schwarer, A.
Hahn, C.
Hughes, T.
White, D.
Ross, D.
Citation: The Journal of molecular diagnostics : JMD, 2017; 19(5):711-721
Publisher: American Society for Investigative Pathology
Issue Date: 2017
ISSN: 1943-7811
Statement of
Ilaria S. Pagani, Chung H. Kok, Verity A. Saunders, Mark B. Van der Hoek, Susan L. Heatley, Anthony P. Schwarer, Christopher N. Hahn, Timothy P. Hughes, Deborah L. White and David M. Ross
Abstract: Somatic mitochondrial DNA (mtDNA) mutations have been identified in many human cancers, including leukemia. To identify somatic mutations, it is necessary to have a control tissue from the same individual for comparison. When patients with leukemia achieve remission, the remission peripheral blood may be a suitable and easily accessible control tissue, but this approach has not previously been applied to the study of mtDNA mutations. We have developed and validated a next-generation sequencing approach for the identification of leukemia-associated mtDNA mutations in 26 chronic myeloid leukemia patients at diagnosis using either nonhematopoietic or remission blood samples as the control. The entire mt genome was amplified by long-range PCR and sequenced using Illumina technology. Variant caller software was used to detect mtDNA somatic mutations, and an empirically determined threshold of 2% was applied to minimize false-positive results because of sequencing errors. Mutations were called against both nonhematopoietic and remission controls: the overall concordance between the two approaches was 81% (73/90 mutations). Some discordant results were because of the presence of somatic mutations in remission samples, because of either minimal residual disease or nonleukemic hematopoietic clones. This method could be applied to study somatic mtDNA mutations in leukemia patients who achieve minimal residual disease, and in patients with nonhematopoietic cancers who have a matched uninvolved tissue available.
Keywords: Humans; Leukemia; DNA, Mitochondrial; Remission Induction; Reproducibility of Results; Polymerase Chain Reaction; Sequence Analysis, DNA; DNA Mutational Analysis; Mutation; Alleles; High-Throughput Nucleotide Sequencing; Biomarkers, Tumor
Rights: © 2017 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.
RMID: 0030073170
DOI: 10.1016/j.jmoldx.2017.05.009
Appears in Collections:Medicine publications

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