Incremental Yield of CD34-Positive Cell Enrichment for Next-Generation Sequencing-Based Measurable Residual Disease Detection in CD34-Positive Acute Myeloid Leukemia

Next-generation sequencing (NGS)-based measurable residual disease (MRD) detection in AML may be limited by non-leukemic hematopoietic cells in bulk bone marrow samples. To address this limitation, we aimed to evaluate whether CD34-positive cell enrichment improves the detectability of residual variants. Paired MRD analyses were performed using unsorted and CD34-enriched bone marrow samples from patients with CD34-positive AML, tracking leukemia-associated variants identified at diagnosis in follow-up samples. Variant-level changes in variant allele frequency (VAF) and sample-level MRD positivity were assessed. CD34 enrichment significantly increased variant-level VAFs (P <0.0001), particularly for low-frequency variants. MRD detection increased by 12.7% (7/55 samples), although overall MRD positivity increased from 27.3% to 36.4% without statistical significance (P =0.18). CD34-positive cell enrichment may improve the detectability of low-frequency variants and provide additional value in selected cases with low disease burden.