Department of Hematology, University of Schleswig-Holstein, Campus Kiel, Langer Segen 8-10, 24105, Kiel, Germany.
CLIP-Childhood Leukaemia Investigation Prague, Department of Paediatric Haematology and Oncology, Second Faculty of Medicine, Charles University, University Hospital Motol, Prague, Czech Republic.
Mol Diagn Ther. 2017 Oct;21(5):481-492. doi: 10.1007/s40291-017-0277-9.
Minimal residual disease (MRD) is the most important independent prognostic factor in acute lymphoblastic leukemia (ALL). Since it has been implemented into in treatment stratification strategies, cure rates have improved significantly for all age groups. Real time quantitative (RQ)-PCR of clonal immunoglobulin and T-cell receptor gene rearrangements using allele-specific primers is currently regarded as the gold standard for MRD analysis in ALL, as it is not only highly sensitive and specific but also provides accurate MRD quantification. Following recent advances in next-generation sequencing (NGS), much attention has been devoted to the development of NGS-based MRD assays. This new technique can enhance sensitivity provided that sufficient numbers of cells are analyzed. Recent reports have shown that NGS-MRD also tends to be more specific for relapse prediction than RQ-PCR. In addition, NGS provides information on the physiological B- and T-cell repertoire during and after treatment, which has been shown to be prognostically relevant. However, before implementation of NGS-MRD detection in clinical practice, several issues must be addressed and the whole workflow needs to be standardized, including not only the analytical phase (spike-in calibrators, quality controls) but also the pre-analytical (e.g. sample preparation) and the post-analytical phases (e.g. bioinformatics pipeline, guidelines for correct data interpretation). These topics are currently addressed by a European network, the EuroClonality-NGS Consortium. In conclusion, NGS is a promising tool for MRD detection with the potential to overcome most of the limitations of RQ-PCR and to become the new gold standard for MRD detection in ALL.
微小残留病 (MRD) 是急性淋巴细胞白血病 (ALL) 最重要的独立预后因素。自从它被纳入治疗分层策略以来，所有年龄段的治愈率都有了显著提高。使用等位基因特异性引物的克隆免疫球蛋白和 T 细胞受体基因重排的实时定量 (RQ)-PCR 目前被认为是 ALL 中 MRD 分析的金标准，因为它不仅高度敏感和特异性，而且还提供了准确的 MRD 定量。随着新一代测序 (NGS) 的最新进展，人们越来越关注基于 NGS 的 MRD 检测方法的开发。这种新技术可以提高灵敏度，只要分析足够数量的细胞即可。最近的报告表明，与 RQ-PCR 相比，NGS-MRD 也更倾向于用于预测复发。此外，NGS 提供了治疗期间和治疗后生理 B 细胞和 T 细胞库的信息，这些信息已被证明与预后相关。然而，在将 NGS-MRD 检测应用于临床实践之前，必须解决几个问题并标准化整个工作流程，包括不仅包括分析阶段（加标校准物、质量控制），还包括分析前阶段（例如样本制备）和分析后阶段（例如生物信息学管道、正确数据解释的指南）。这些主题目前由一个欧洲网络，即 EuroClonality-NGS 联盟来处理。总之，NGS 是一种很有前途的 MRD 检测工具，具有克服 RQ-PCR 大多数局限性的潜力，并可能成为 ALL 中 MRD 检测的新标准。
