Webinar: How NGS Can Rapidly Deliver Key Insights for Myeloid Neoplasms

Webinar summary

In this webinar, Dr. Bevan Tandon, Director of Hematopathology and Molecular Pathology at Pathline Labs, discusses the application of next-generation sequencing (NGS) for genomic characterization and minimal residual disease (MRD) detection in myeloid neoplasms.

The presentation highlights the use of targeted multi-gene sequencing panels to identify genomic alterations, assess clonal architecture, and enable high-sensitivity detection of low-frequency variants across various myeloid neoplasms.

Study overview

Myeloid neoplasms represent genetically heterogeneous systems characterized by diverse genomic alterations, including single nucleotide variants (SNVs), insertions and deletions (indels), structural rearrangements, and copy number variations.

Conventional testing algorithms employ multiple independent methodologies, including morphology, immunophenotyping, cytogenetics, fluorescence-based hybridization, and PCR. These approaches are typically iterative, limited in genomic breadth, and variable in analytical sensitivity.

Targeted NGS assays enable multiplexed interrogation of numerous genes within a single test. This study evaluates the Oncomine Myeloid Assay GX v2 on the Ion Torrent Genexus Integrated Sequencer, an NGS platform with automated library preparation, sequencing, and bioinformatic analysis in one simple workflow to enable rapid and comprehensive genomic profiling.

The study also assesses the Oncomine Myeloid MRD Assays (RUO) on the GeneStudio S5 System to demonstrate the high-level of sensitivity that can be achieved using error-corrected NGS for MRD analysis.

Analytical verification included assessment of accuracy, reproducibility, sensitivity, and concordance using reference materials and comparative sequencing platforms. Representative datasets were analyzed across multiple myeloid neoplasms, including acute myeloid leukemia, myelodysplastic syndromes, and myeloproliferative neoplasms.

Key findings

Limitations of conventional analytical methods

• Sequential single-target assays introduce workflow complexity and limit comprehensive genomic coverage.
• PCR and Sanger sequencing approaches exhibit reduced sensitivity relative to NGS.
• Iterative testing strategies increase total processing time and may fail to capture co-occurring genomic alterations.

Analytical performance of targeted NGS using Oncomine Myeloid Assay GX v2

• Enabled simultaneous detection of multiple genomic variants across a defined gene set.
• Demonstrated high analytical accuracy and reproducibility (>98%) with strong concordance across sequencing platforms.
• Variant detection sensitivity was established at approximately 4–5% variant allele frequency, exceeding that of conventional sequencing approaches.
• Identified low-frequency and co-occurring genomic variants that were not detectable by conventional methods.
• Multiplex analysis enabled characterization of complex mutational patterns and co-mutation profiles across the various myeloid diseases.

Workflow integration and throughput on the Genexus Integrated Sequencer

• Automated NGS workflows enabled rapid sample-to-data generation within approximately 24–48 hours.
• Integration of sequencing and bioinformatic pipelines reduced manual intervention and streamlined data interpretation.
• Consolidation of multiple analytical steps improved operational efficiency and reproducibility.

High-sensitivity MRD analysis using Oncomine Myeloid MRD Assays (RUO)

• Use of unique molecular identifiers (UMIs) enabled error correction and increased analytical sensitivity.
• Detection thresholds approached ~0.1% variant allele frequency, with potential sensitivity to lower-frequency variants under optimized conditions.
• Multiplex tracking of genomic variants enabled longitudinal analysis of clonal persistence and subclonal dynamics.
• NGS-based approaches demonstrated greater sensitivity for detection of low-frequency variants compared to immunophenotypic and targeted PCR methods.

Targeted NGS provides a high-resolution, multiplexed framework for genomic analysis of myeloid neoplasms. The approach enables comprehensive detection of diverse genomic alterations, improved sensitivity for low-frequency variants, and enhanced characterization of clonal architecture.

Integration of molecular barcoding strategies further extends analytical capability for detection of residual genomic signal and monitoring of clonal dynamics. These findings support the utility of NGS as a foundational technology for genomic research in myeloid neoplasms and for advancing understanding of mutation persistence and evolution.