Next-Generation Sequencing for Minimal Residual Disease Detection in AML: Current Technologies and Clinical Implications.

Abstract

Minimal residual disease (MRD) has become a significant predictor of relapse and survival in acute myeloid leukemia (AML), indicating the extent of remission beyond traditional morphological evaluation. Although multicolor flow cytometry and quantitative PCR are essential methodologies in minimal residual disease identification, both are constrained by immunophenotypic variability, the necessity for stable molecular targets, and limited sensitivity. Advancements in next-generation sequencing (NGS) have revolutionized the minimal residual disease (MRD) field by enabling highly sensitive, mutation-driven identification of leukemic clones across a broad genomic landscape. Contemporary error-suppressed next-generation sequencing techniques—such as unique molecular identifiers, duplex sequencing, and single-molecule molecular inversion probes—have enhanced analytical sensitivity to the 10⁻⁵ to 10⁻⁶ range, enabling the detection of ultra-low-frequency variations with greater specificity. These techniques improve clinical risk classification, refine prognostication within genetically defined AML subtypes, and guide therapeutic options, including post-remission therapy, targeted inhibition, and the timing and intensity of allogeneic stem cell transplantation. Innovative applications, such as single-cell sequencing, cell-free DNA studies, and integrative multi-omic MRD evaluation, enhance the capabilities of genomics-based monitoring. Nonetheless, obstacles remain, such as differentiating cancer mutations from clonal hematopoiesis, standardizing analytical pipelines, establishing clinically relevant thresholds, and incorporating NGS MRD into standardized treatment protocols. This review encapsulates contemporary NGS methods for AML MRD diagnosis, assesses their clinical ramifications and constraints, and suggests future pathways necessary for comprehensive clinical integration. With advancements in the area, NGS-based MRD is set to become a pivotal element of precision-guided AML control.

Affiliated Institutions

• Fort Hays State University US
• Kingston University GB
• University of Vermont Medical Center US
• University of North Carolina at Greensboro US
• University of Nigeria NG
• Nnamdi Azikiwe University Teaching Hospital NG
• University of Benin NG

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