Table 3.

Pros and cons of methods used to detect MRD in AML

TechnologySensitivityProsCons
MFC∼10−4 to 10−5Wide applicability (>90%)Challenging and somewhat subjective interpretation requires experienced pathologist
Relatively quick (results ≤1 d)Sensitivity dependent on antibody panel used
Single result interpretableLimited harmonization and standardization across laboratories
High specificity when using defined LAIPLeukemic 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 10−5Relatively easy to performLimited standardization
SensitiveError rate leads to low sensitivity of mutated sequences
Applicable to specific subgroupsMutated 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 10−5Wide applicabilityResults may take multiple days
May be run by any certified laboratory with RT-qPCR capacityExpensive (computationally demanding and time-consuming)
High sensitivity (≥MFC)Requires high-level expertise
Well standardizedRequires setting of threshold limits
Quality assurance routinely incorporatedInterpretation 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 10−2Superior 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 analysisNAMore common in routine clinical practiceLabor-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.