Department of Hematology, Hemostasis, Oncology, and Stem Cell Transplantation, Hannover Medical School, Hannover, Germany.
Genes Chromosomes Cancer. 2012 Jul;51(7):689-95. doi: 10.1002/gcc.21955. Epub 2012 Mar 27.
Systematic assessment of minimal residual disease (MRD) in acute myeloid leukemia (AML) patients has been hampered by lack of a reliable, uniform MRD marker applicable to all patients. We evaluated next-generation sequencing (NGS) for MRD assessment in AML patients (n = 80 samples). The ability of NGS technologies to generate thousands of clonal sequences makes it possible to determine the allelic ratio of sequence variants. Using NGS, we were able to determine the allelic ratio of different FLT3-internal tandem duplication (ITD) clones within one patient sample, in addition to resolution of FLT3-ITD insertion site, length, and sequence in a single analysis. Furthermore, NGS allowed us to study emergence of clonal dominance. Parallel assessment of MRD by NGS and quantitative real-time polymerase chain reaction in NPM1 mutated patients was concordant in 95% of analyzed samples (n = 38). The frequency of mutated alleles was linearly quantified by NGS. As NGS sensitivity is scalable depending on sequence coverage, it reflects a highly flexible and reliable tool to assess MRD in leukemia patients.
系统评估急性髓细胞白血病（AML）患者的微小残留病（MRD）一直受到缺乏适用于所有患者的可靠、统一的 MRD 标志物的阻碍。我们评估了下一代测序（NGS）在 AML 患者（n=80 个样本）中的 MRD 评估能力。NGS 技术能够生成数千个克隆序列，这使得确定序列变异的等位基因比成为可能。使用 NGS，我们能够确定一个患者样本中不同 FLT3 内部串联重复（ITD）克隆的等位基因比，此外还可以在单次分析中确定 FLT3-ITD 插入位点、长度和序列。此外，NGS 还允许我们研究克隆优势的出现。在 NPM1 突变患者中，NGS 和实时定量聚合酶链反应（qPCR）平行评估 MRD 的结果在 95%的分析样本（n=38）中是一致的。NGS 可线性定量突变等位基因的频率。由于 NGS 的灵敏度可根据序列覆盖度进行扩展，因此它反映了一种非常灵活和可靠的工具，可用于评估白血病患者的 MRD。
