The research-use-only assay combines two key genetic markers for acute myeloid leukemia in a single test, potentially reducing the need for separate testing workflows and reference lab send-outs.

Pillar Biosciences has launched the MRDTect NPM1 and FLT3 ITD Panel, a research-use-only, kitted next-generation sequencing (NGS) assay designed for measurable residual disease (MRD) detection in acute myeloid leukemia. The assay marks the company’s entry into MRD testing, uniting NPM1 exon 12 mutation and FLT3 internal tandem duplication (ITD) detection in a single-tube, DNA-based workflow capable of detecting variants down to a 0.005% variant allele frequency (VAF).

The launch aims to help clinical laboratories bring ultrasensitive molecular monitoring in-house using existing benchtop sequencing infrastructure, offering an alternative to centralized reference laboratory send-outs.

MRD serves as an essential prognostic marker in acute myeloid leukemia management to assess therapeutic response and detect impending relapse earlier than standard morphology. The 2025 European LeukemiaNet (ELN) MRD recommendations advise evaluating leukemia-specific molecular markers at limits of detection between 10⁻⁴ and 10⁻⁵, while introducing ultrahigh-sensitivity NGS for FLT3-ITD-positive disease. Currently, laboratories frequently navigate a fragmented workflow: outsourcing samples or running separate internal assays, such as RNA-based quantitative reverse transcription PCR (qRT-PCR) for NPM1 and hybrid-capture NGS with long-read sequencing for FLT3-ITD. This division typically increases DNA and RNA input requirements, operational costs, manual labor, and turnaround times.

“MRD is where precision oncology is heading, and laboratories need tools that make ultrasensitive monitoring practical in-house,” says ShiPing Zou, PhD, senior director of product strategy and portfolio management of Pillar Biosciences, in a release. “MRDTect NPM1 and FLT3 ITD Panel is our first step in helping laboratories localize NGS-based MRD testing. It brings two of the most informative [acute myeloid leukemia] markers into one simple, kitted workflow that runs on the sequencers in their own labs, built on the same SLIMamp chemistry our customers already trust.”

Single-Tube Workflow and Analytical Specifications

The MRDTect panel utilizes Pillar’s proprietary Stem-Loop Inhibition-Mediated amplification technology alongside its AI-driven VersaTile Primer Design platform. Both genetic targets are amplified in a single reaction tube and sequenced using standard 2 × 150 bp paired-end sequencing configurations, bypassing the requirement for specialized long-read sequencing platforms.

Key technical specifications of the panel include:

  • Analytical Sensitivity: Demonstrates 100% detection of NPM1 and FLT3-ITD variants at 0.005% VAF (5 × 10⁻⁵) and above, satisfying ELN-recommended detection thresholds, with detections also recorded below that level.
  • Broad Insertion Coverage: Identifies ITD insertions ranging from approximately 3 base pairs to more than 300 base pairs, capturing ultra-long ITD mutations that often elude standard testing approaches.
  • DNA Input and Sample Flexibility: Requires 500 ng to 700 ng of genomic DNA for a target VAF of 10⁻⁴, or 700 ng for a target VAF of 10⁻⁵, isolated from whole blood, peripheral blood mononuclear cells, white blood cells, or bone marrow. Using a single DNA input mitigates RNA pre-analytical variability and conserves limited clinical specimens.
  • Turnaround Efficiency: Features approximately 3.5 hours of hands-on time, under 8.5 hours from DNA extraction to sequencer loading, flexible workflow stopping points, and overall sample-to-result delivery in under 48 hours.
  • Sequencing and Informatics: Requires 1.2 million or more paired-end reads per sample for a 10⁻⁴ target VAF and 8.9 million or more paired-end reads for a 10⁻⁵ target VAF. Data processing is managed by the company’s PiVAT analysis software, which analyzes each sample in 3 to 10 minutes and includes dedicated long-indel calling and automated quality-control metrics.

Multi-Laboratory Evaluation

Pillar characterized the performance of the assay internally and across four external sites, including an academic cancer center and a large commercial reference laboratory. Across more than 200 sequencing libraries prepared from reference standards and clinical specimens, median mapping rates exceeded 99% across all sample types, with approximately 99.2% of aligned reads falling on target alongside balanced coverage across NPM1 and FLT3 regions.

The panel is designated for research use only and is not cleared for use in diagnostic procedures. Pillar plans to expand the MRDTect platform to support localized MRD monitoring across other hematologic malignancies in future releases.

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