Measurable residual disease (MRD) detected by multiparametric flow cytometry (MFC) is associated with unfavorable outcome in patients with AML. A simple, broadly applicable eight-color panel was implemented and analyzed utilizing a hierarchical gating strategy with fixed gates to develop a clear-cut LAIP-based DfN approach. In total, 32 subpopulations with aberrant phenotypes with/without expression of markers of immaturity were monitored in 246 AML patients after completion of induction chemotherapy. Reference values were established utilizing 90 leukemia-free controls. Overall, 73% of patients achieved a response by cytomorphology. In responders, the overall survival was shorter for MRD^pos patients (HR 3.8, p = 0.006). Overall survival of MRD^neg non-responders was comparable to MRD^neg responders. The inter-rater-reliability for MRD detection was high with a Krippendorffs α of 0.860. The mean time requirement for MRD analyses at follow-up was very short with 04:31 minutes. The proposed one-tube MFC approach for detection of MRD allows a high level of standardization leading to a promising inter-observer-reliability with a fast turnover. MRD defined by this strategy provides relevant prognostic information and establishes aberrancies outside of cell populations with markers of immaturity as an independent risk feature. Our results imply that this strategy may provide the base for multicentric immunophenotypic MRD assessment.

多参数流式细胞术 (MFC) 检测到的可测量残留病灶 (MRD) 与 AML 患者的不良预后相关。利用具有固定门的分层选通策略来实现和分析简单、广泛适用的八色面板，以开发清晰的基于 LAIP 的 DfN 方法。总共，在完成诱导化疗后，对 246 名 AML 患者中的 32 个具有异常表型（有/无不成熟标志物表达）的亚群进行了监测。利用 90 个无白血病对照建立参考值。总体而言，73% 的患者获得了细胞形态学缓解。^{在应答者中，MRD pos}患者的总生存期较短（HR 3.8，p  = 0.006）。MRD^{阴性无反应者的总生存率与 MRD阴性}反应者相当。MRD 检测的评估者间可靠性很高，Krippendorffs α 为 0.860。随访时 MRD 分析的平均时间要求非常短，为 04:31 分钟。所提出的用于检测 MRD 的单管 MFC 方法允许高水平的标准化，从而带来有希望的观察者间可靠性和快速周转。该策略定义的 MRD 提供了相关的预后信息，并确定了细胞群之外的异常情况，并将不成熟标记作为独立的风险特征。我们的结果表明该策略可以为多中心免疫表型 MRD 评估提供基础。