MRD detection in AML: remaining key issues
| Detection method . | Issue . |
|---|---|
| All | Definition of MRD positivity/negativity |
| Differences in source of material (bone marrow vs peripheral blood) | |
| For bone marrow: hemodilution | |
| Variation in timing/frequency of MRD sampling | |
| Regulatory approval/validation of assay | |
| Insufficient assay sensitivity | |
| Disease evolution with change in targets suitable for MRD detection | |
| Requirement of expertise for data interpretation | |
| Sample degradation | |
| Molecular | Choice of target(s), target specificity |
| Quality of cDNA synthesis | |
| Efficacy of PCR amplification | |
| Insufficient primer specificity | |
| Sensitivity of target gene overexpression limited by normal tissue expression | |
| Target stability | |
| Data normalization; choice of housekeeping gene | |
| Contamination | |
| Flow cytometry | Choice of antigens and antibody panels |
| Lack of immunophenotypic abnormalities | |
| Lack of diagnostic specimen to determine immunophenotypic abnormalities sufficient for MRD detection | |
| Choice of analysis strategy for MRD detection (diagnostic leukemia-associated immunophenotypes vs “different-from-normal” analysis) | |
| Lack of automatic analysis algorithms |
| Detection method . | Issue . |
|---|---|
| All | Definition of MRD positivity/negativity |
| Differences in source of material (bone marrow vs peripheral blood) | |
| For bone marrow: hemodilution | |
| Variation in timing/frequency of MRD sampling | |
| Regulatory approval/validation of assay | |
| Insufficient assay sensitivity | |
| Disease evolution with change in targets suitable for MRD detection | |
| Requirement of expertise for data interpretation | |
| Sample degradation | |
| Molecular | Choice of target(s), target specificity |
| Quality of cDNA synthesis | |
| Efficacy of PCR amplification | |
| Insufficient primer specificity | |
| Sensitivity of target gene overexpression limited by normal tissue expression | |
| Target stability | |
| Data normalization; choice of housekeeping gene | |
| Contamination | |
| Flow cytometry | Choice of antigens and antibody panels |
| Lack of immunophenotypic abnormalities | |
| Lack of diagnostic specimen to determine immunophenotypic abnormalities sufficient for MRD detection | |
| Choice of analysis strategy for MRD detection (diagnostic leukemia-associated immunophenotypes vs “different-from-normal” analysis) | |
| Lack of automatic analysis algorithms |
cDNA, complementary DNA.
Key issues for MRD detection in AML have been previously highlighted by several investigators.1-4,36,40,41,43,44,47-51