^oncoReveal®
Myeloid v2 Tech Note 

oncoReveal® Myeloid v2 Panel 
Data characterization and panel performance 

Introduction
Myeloid malignancies comprise a diverse group of hematologic neoplasms, including acute myeloid leukemia (AML), myelodysplastic syndromes (MDS), myeloproliferative neoplasms (MPNs), chronic myelomonocytic leukemia (CMML), and related disorders. These diseases are driven by a broad spectrum of clinically relevant genomic alterations that support disease classification, prognostic risk stratification, therapeutic selection, and measurable residual disease assessment. Current clinical guidelines from organizations, such as the WHO, NCCN, and AMP/CAP, increasingly recommend comprehensive molecular profiling across multiple genes and variant classes. As a result, laboratories require genomic assays that can simultaneously detect single nucleotide variants (SNVs), insertions/deletions (indels), and complex alterations such as FLT3 internal tandem duplications (ITDs). These assays must also maintain reliable coverage across technically challenging, GC-rich regions such as CEBPA. Targeted NGS meets this need by consolidating profiling into one assay, reducing specimen use, streamlining workflows, and reducing turnaround time.

The oncoReveal® Myeloid v2 Panel addresses these requirements using Pillar Biosciences’ SLIMamp® multiplex PCR technology. It interrogates 80 myeloid-associated genes, with full CDS for 18 genes, in a single-tube, same-day library preparation workflow, with robust detection of SNVs, indels, and FLT3 ITDs and uniform coverage of challenging regions such as high GC content regions in CEBPA. This application note characterizes sequencing performance, coverage uniformity, analytical sensitivity, variant allele frequency (VAF) accuracy, and FLT3 ITD detection across reference materials and clinical specimens.

Materials and Methods
Panel design and workflow

The oncoReveal Myeloid v2 Panel was developed by expanding and updating the content of the original oncoReveal Myeloid assay. Target selection was guided by review of major myeloid guidelines, including WHO 2016 and 2022, ICC 2022, ELN 2022, and NCCN recommendations. The resulting panel comprises 874 amplicons covering 80 genes associated with myeloid malignancies, including full coding sequence coverage of 18 genes and targeted hotspot coverage of 62 genes (Table 1). The panel is designed to detect SNVs and indels down to ~1% VAF, and FLT3 ITDs from 3 bp to ≥300 bp (upper bound ITD length not estimated) down to ~0.3%.