QIAact Myeloid DNA UMI Panel

• Targeting 25 genes of known significance to myeloid malignancies
• High sensitivity for detection of key variants, enabled by digital sequencing using UMIs
• Detection of challenging mutations including FLT3 ITDs, CALR deletions and CEBPA mutations
• Verified as part of our complete Sample to Insight workflow
• Integrated bioinformatics interpretation and reporting with QCI

The QIAact Myeloid DNA UMI Panel in combination with the QIAGEN GeneReader NGS System provides a single integrated solution to simultaneously test for about 9000 variants in 25 genes with reported relevance to clonal myeloid malignancy. The incorporation of unique molecular index (UMI) technology enables detection of low frequency variants, including those below the 1% variant allele frequency (VAF) such as mutations within JAK2 and KIT genes (for the other genes, the panel supports mutation detection with a VAF of 5%). Accurate reporting of usually challenging-to-detect mutations including the large CALR Type 1 (52 bp deletion), FLT3 ITDs (with identified insertion site), and GC-rich sequences, such as in the CEBPA gene, are also possible due to this digital sequencing approach paired with optimized bioinformatics.

This Sample to Insight solution can detect pathogenic and actionable variants from both blood and bone marrow. Integrated as part of the complete GeneReader NGS System workflow, the panel offers a complete assay, including QCI Analyze and Interpret for seamless variant reporting based on the latest AMP/ASCO/CAP guidelines for somatic variant classification.

DNA sequencing is a useful tool to detect genetic variations, including somatic mutations, single nucleotide variants (SNVs) and insertions and deletions (InDels). Target enrichment technology enables next-generation sequencing (NGS) platform users to sequence specific regions of interest instead of the entire genome, effectively increasing sequencing depth and throughput with lower cost. Existing target enrichment methods, library preparation and sequencing steps all utilize enzymes and amplification processes, which introduce substantial bias and artefacts. The resultant background artefactual errors greatly limit the detection of true low-frequency variants in heterogeneous cancer samples.

The QIAact Myeloid DNA UMI Panel integrates unique molecular index (UMI) technology into a gene-specific, primer-based target enrichment process, enabling sensitive variant detection of targeted genomic regions by NGS on the GeneReader NGS System.

The QIAact Myeloid DNA UMI Panel has been optimized in combination with a specially formulated enrichment chemistry to achieve highly efficient enrichment on both regular and GC-rich regions at high multiplex levels.

The QIAact Myeloid DNA UMI Panel is provided as two primer mix tubes, with approximately 400 primers per tube. The QIAact Myeloid DNA UMI Panel is designed to enrich specific target regions in select genes (ASXL1, CALR, CBL, CEBPα, CSF3R, DNMT3A, EZH2, FLT3, IDH1, IDH2, JAK2, KIT, KRAS, MPL, NPM1, NRAS, RUNX1, SETBP1, SF3B1, SH2B3, SRSF2, TET2, TP53, U2AF1, ZRSR2) using 40 ng of DNA.

Genomic DNA samples are first fragmented, end-repaired and A-tailed using a single, controlled multi-enzyme reaction. The prepared DNA fragments are then ligated at their 5’ ends to a GeneReader specific adapter containing a unique molecular index (UMI) and a nine (9) base-pair (bp) sample-specific bar code.

Ligated DNA molecules are subject to limited cycles of target enrichment PCR, with one gene-specific primer targeting a region and one universal forward primer complimentary to an adapter sequence. This reaction ensures that intended targets and UMIs are enriched sufficiently to be represented in the final library. A universal PCR with GeneReader specific sequences is then carried out to amplify the targets and complete the library.

Once the library is sequenced, results can be analyzed using the QIAact Myeloid DNA UMI Panel workflow, which will automatically perform all steps necessary to generate a DNA sequence variant report from your raw NGS data. All detected variants can be further interpreted by QIAGEN Clinical Insight (QCI) analysis.

For targeted enrichment prior to next-generation sequencing (NGS) using the QIAGEN GeneReader instrument