Pros and cons of methods used to detect MRD in AML
| Technology . | Sensitivity . | Pros . | Cons . |
|---|---|---|---|
| MFC | ∼10−4 to 10−5 | Wide applicability (>90%) | Challenging and somewhat subjective interpretation requires experienced pathologist |
| Relatively quick (results ≤1 d) | Sensitivity dependent on antibody panel used | ||
| Single result interpretable | Limited harmonization and standardization across laboratories | ||
| High specificity when using defined LAIP | Leukemic phenotype not necessarily stable over time (eg, initial LAIP may not identify subclones leading to relapse) | ||
| Can detect cells with leukemia-stem cell phenotype | |||
| Can distinguish between live and dead cells | |||
| Ease of data storage | |||
| Provides information about whole sample cellularity | |||
| NGS | ∼10−3 to 10−5 | Relatively easy to perform | Limited standardization |
| Sensitive | Error rate leads to low sensitivity of mutated sequences | ||
| Applicable to specific subgroups | Mutated genes can be detected in healthy people without hematologic abnormalities | ||
| Persistence of some genetic abnormalities in patients in long-term remission | |||
| Risk of contamination | |||
| RT-qPCR | ∼10−3 to 10−5 | Wide applicability | Results may take multiple days |
| May be run by any certified laboratory with RT-qPCR capacity | Expensive (computationally demanding and time-consuming) | ||
| High sensitivity (≥MFC) | Requires high-level expertise | ||
| Well standardized | Requires setting of threshold limits | ||
| Quality assurance routinely incorporated | Interpretation often requires trend of results | ||
| Different mutations have different biological consequences in AML | |||
| Molecular targets applicable to only ∼50% of all AML cases and <35% in older patents | |||
| FISH | ∼1 to 10−2 | Superior to PCR-based assays for detection of numeric cytogenetic abnormalities (gains and losses of whole chromosomes or deletions/duplications) | Considerably less sensitive than PCR or MFC |
| Not useful for patients with normal karyotype | |||
| Quality-assured probes are expensive; technique is labor intensive | |||
| Chromosome banding analysis | NA | More common in routine clinical practice | Labor-intensive, comparatively costly, low-throughput technique reliant on highly trained technical staff |
| Evaluates the dividing proportion of cells | ∼10× less sensitive than FISH | ||
| Not useful for patients with normal karyotype |
| Technology . | Sensitivity . | Pros . | Cons . |
|---|---|---|---|
| MFC | ∼10−4 to 10−5 | Wide applicability (>90%) | Challenging and somewhat subjective interpretation requires experienced pathologist |
| Relatively quick (results ≤1 d) | Sensitivity dependent on antibody panel used | ||
| Single result interpretable | Limited harmonization and standardization across laboratories | ||
| High specificity when using defined LAIP | Leukemic phenotype not necessarily stable over time (eg, initial LAIP may not identify subclones leading to relapse) | ||
| Can detect cells with leukemia-stem cell phenotype | |||
| Can distinguish between live and dead cells | |||
| Ease of data storage | |||
| Provides information about whole sample cellularity | |||
| NGS | ∼10−3 to 10−5 | Relatively easy to perform | Limited standardization |
| Sensitive | Error rate leads to low sensitivity of mutated sequences | ||
| Applicable to specific subgroups | Mutated genes can be detected in healthy people without hematologic abnormalities | ||
| Persistence of some genetic abnormalities in patients in long-term remission | |||
| Risk of contamination | |||
| RT-qPCR | ∼10−3 to 10−5 | Wide applicability | Results may take multiple days |
| May be run by any certified laboratory with RT-qPCR capacity | Expensive (computationally demanding and time-consuming) | ||
| High sensitivity (≥MFC) | Requires high-level expertise | ||
| Well standardized | Requires setting of threshold limits | ||
| Quality assurance routinely incorporated | Interpretation often requires trend of results | ||
| Different mutations have different biological consequences in AML | |||
| Molecular targets applicable to only ∼50% of all AML cases and <35% in older patents | |||
| FISH | ∼1 to 10−2 | Superior to PCR-based assays for detection of numeric cytogenetic abnormalities (gains and losses of whole chromosomes or deletions/duplications) | Considerably less sensitive than PCR or MFC |
| Not useful for patients with normal karyotype | |||
| Quality-assured probes are expensive; technique is labor intensive | |||
| Chromosome banding analysis | NA | More common in routine clinical practice | Labor-intensive, comparatively costly, low-throughput technique reliant on highly trained technical staff |
| Evaluates the dividing proportion of cells | ∼10× less sensitive than FISH | ||
| Not useful for patients with normal karyotype |
FISH, fluorescence in situ hybridization; NA, not available. QA, quality assurance.