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. 2020 Jun 3;9(6):1714. doi: 10.3390/jcm9061714

Table 2.

Comparison of strategies used for MRD detection using multiparametric flow cytometry (MFC) and their clinical evidence.

Reference MFC Method Methodology/Surface Markers Used Time Point of Assessment Evidence-Outcomes
[42] MFC-LAIP Limit of detection 0.1% After induction therapy

  • MRDneg: median RFS >47 months; 4-year RFS 52%

  • MRDpos: median RFS 8.6 months; 4-year RFS 23%
[50] MFC-LAIP combined with qPCR 8 color MFC assay After consolidation

  • Both neg: 2-year OS 89% and DFS 69%

  • MFCpos/PCRneg or MFCneg/PCR pos: 2-year OS 88–89%; DFS 65–76%

  • Both pos: 2-year OS 55%, DFS 22%
[26] MFC-LAIP combined with NGS Limit of detection 0.1% After induction therapy

  • Both pos: 4-year relapse rate 73.3%

  • NGSpos/MFCneg: 4-year relapse 52.3%

  • NGSneg/MFCpos: 4-year relapse 49.8%

  • Both neg: 4-year relapse rate 26.7%
[49] LAIP at time of bone marrow aplasia (day 16–18) 31 surface markers, multiple LAIPs identified for each patient
Limit of detection 0.15%		 Day 16–18 of induction therapy

  • MRDneg: 5-year RFS 43%

  • MRDpos: 5-year RFS 16%
[10] MFC-DfN 10 color assay
3 tubes, 1 million events per tube.
Limit of detection 0.1%		 MRD assessment pre-alloSCT and outcomes post-transplant

  • MRDneg: 3-year OS >70%; relapse risk 20–25%

  • MRDpos: 3-year OS 25%, relapse risk 70%
[51] MFC-DfN 10 color MFC assay
Surface markers: CD4, CD5, CD7, CD13, CD14, CD15, CD16, CD19, CD33, CD34, CD38, CD45, CD56, CD64, CD71, CD117, CD123, HLA-DR.
Any measurable MRD considered positive		 Pre-alloSCT and post-alloSCT (day +28)

  • MRDpos pre-alloSCT and neg post: 3-year OS 29% and RFS 18%

  • MRDpos both timepoints: 3-year OS 19%, 3-year RFS 14%

  • MRDneg at both time points: OS 76%, RFS 71%
[29] MFC-DfN separately and combined with NGS 10 color MFC assay
Limit of detection 0.1%		 Pre-alloSCT

  • MFC–MRDpos: 18-month relapse incidence 37%; OS 48%

  • MFC–MRDneg: 18-month relapse 9%; OS 73%

  • Both MFC and NGS pos: 18-month relapse 51%; OS 51%

  • Both MFC and NGS neg: 18-month relapse 8%; OS 78%

  • MFCneg/NGSpos or MFCpos/NGSneg: 18-month relapse 17%; OS 44%
[52] Combined MFC-LAIP and LSC Limit of detection 0.1%
LSCs defined as CD34+/CD38− 		After second induction

  • Both neg: 3-year OS 66%, CIR 35%

  • MRDpos and LSCneg: OS 68%, CIR 43%

  • MRDneg and LSCpos: OS 53%, CIR 53%

  • Both pos: OS 0%, CIR 10%
[53] Novel leukemia-specific markers CD9, CD18, CD25, CD32, CD44, CD47, CD52, CD54, CD59, CD64, CD68, CD86, CD93, CD96, CD97, CD99, CD123, CD200, CD300a/c, CD366, CD371, CX3CR1 At diagnosis (to identify novel markers), compared to healthy donors and relapsed AML patients.

  • Improved sensitivity (1 in 105)

  • Clinical outcomes yet to be studied

LAIP = leukaemia-associated immunophenotype; RFS = relapse-free survival; qPCR = real time quantitative polymerase chain reaction; OS = overall survival; DFS = disease-free survival; NGS = next generation sequencing; DfN = different from normal; alloSCT = allogeneic stem cell transplant; LSC = leukaemic stem cells; CIR = cumulative incidence of relapse; AML = acute myeloid leukaemia.