Methods for detection of MRD in AML
| . | Method . | Target . | Sensitivity . | Applicable in % of AML . | Turn-around time (days) . | Limitations/problems . |
|---|---|---|---|---|---|---|
| Established | Multi-parameter flow cytometry (MFC) | Leukemia-associated immunophenotype (LAIP) or different from normal (DfN) | 10−3 to 10−4 | 85-90 | 2 | Less sensitive, more subjective analysis |
| Established | Real-time quantitative PCR (RT-qPCR) | Robust data: NPM1, CBFB::MYH11, RUNX1::RUNX1T1 Less validated: KMT2A::MLLT3, DEK::NUP214, BCR::ABL1, WT1 | 10−4 to 10−5 | 40-50* | 3-5 | Limited applicability |
| Exploratory | Next-generation sequencing (NGS)†,‡ | Potentially any somatic mutation† | 10−2 to 10−4 | ∼100 | 5-10 | Less sensitive, costly, technically challenging |
| Exploratory | Digital PCR (dPCR) | Specific targeted mutations | 10−3 to 10−4 | ∼70 | 3-5 | Specific assay necessary for every mutation, limited sensitivity |
| . | Method . | Target . | Sensitivity . | Applicable in % of AML . | Turn-around time (days) . | Limitations/problems . |
|---|---|---|---|---|---|---|
| Established | Multi-parameter flow cytometry (MFC) | Leukemia-associated immunophenotype (LAIP) or different from normal (DfN) | 10−3 to 10−4 | 85-90 | 2 | Less sensitive, more subjective analysis |
| Established | Real-time quantitative PCR (RT-qPCR) | Robust data: NPM1, CBFB::MYH11, RUNX1::RUNX1T1 Less validated: KMT2A::MLLT3, DEK::NUP214, BCR::ABL1, WT1 | 10−4 to 10−5 | 40-50* | 3-5 | Limited applicability |
| Exploratory | Next-generation sequencing (NGS)†,‡ | Potentially any somatic mutation† | 10−2 to 10−4 | ∼100 | 5-10 | Less sensitive, costly, technically challenging |
| Exploratory | Digital PCR (dPCR) | Specific targeted mutations | 10−3 to 10−4 | ∼70 | 3-5 | Specific assay necessary for every mutation, limited sensitivity |
Less frequent in elderly patients with AML.
The NGS-MRD threshold has not been defined for individual mutations; NGS-MRD positivity is provisionally defined as ≥ 0.1% variant allele frequency, excluding mutations related to clonal hematopoiesis and germline mutations.
Common gene mutations consistent with pre-malignant clonal hematopoiesis such as DNMT3A, TET2, and AXSL1 excluded; further study is required to determine which mutations are truly indicative of residual AML and not clonal hematopoiesis.