Contributions
Abstract: PB1667
Type: Publication Only
Background
Myeloid malignancies comprise many different subtypes amongst which myeloproliferative neoplasms (MPN), myelodysplastic syndrome (MDS), myelodysplastic/myeloproliferative neoplasms (MDS/MPN) and acute myeloid leukemia (AML), are associated with somatic mutations acquired in around 20-30 key genes. To detect these relevant myeloid malignancy-related genetic variants, a targeted enrichment approach coupled to NGS analysis nicely complement the individual biomarker tests to provide a comprehensive mutational overview.
The QIAact Myeloid DNA UMI Panel in combination with the QIAGEN GeneReader™ NGS System provides an integrated solution to simultaneously interrogate many candidate genes for actionable mutations, from as little as 40 ng input DNA, shortening test time and enabling simplification of lab operations.
The QIAact Myeloid DNA UMI Panel is a 25-gene targeted sequencing panel for markers of known significance to clonal myeloid malignancies, allowing reliable and sensitive detection of single nucleotide variants (SNV) and large Insertion/Deletion (InDel) mutations.
Aims
To assess the QIAact Myeloid DNA UMI Panel performance in combination with the QIAGEN GeneReader NGS System.
Methods
Following the advice from key opinion leaders in the field of blood cancer disorders, the QIAact Myeloid DNA UMI Panel has been designed to detect relevant mutations throughout the most informative genes linked to myeloid disease. A key feature of the panel is the addition of a unique molecular index (UMI) to tag individual molecules prior to target enrichment by PCR. This enables sequencing and PCR bias correction, allowing sensitive detection of mutations (e.g. below 1% MAF for JAK2 (exon 12, 13, 14 & 15) & KIT (exon 8, 9, 10, 11 & 17)).
To assess the assay performance, control samples and blood and bone marrow samples were used. Following target enrichment, libraries were sequenced on the GeneReader™ NGS system and mutations identified using the QIAGEN Clinical Insight (QCI™) Analyze software suite, adjusted to specifically support variant calling in this assay.
Results
Complex mutations like the 52 bp deletion CALR type 1 variant, FLT3 ITDs up to 121 bp insertion and NPM1 insertions were efficiently identified. Limit of detection testing using the power of UMIs demonstrated uniform amplification and sequencing coverage to consistently detect mutations with a 1% MAF and below for JAK2 and KIT (KIT D816V variant can be detected at 0.2%), and 5% for all the other genes covered by the panel. Overall mutational variants were correctly called, resulting in >99% concordance with an alternative testing technology.
Conclusion
The QIAact Myeloid DNA UMI Panel in combination with the QIAGEN GeneReader NGS System offer a fully integrated sample to insight solution. The optimized chemistry allows superior analytical sensitivity in accurately detecting highly relevant genetic alterations for myeloid malignancy research.
Session topic: 3. Acute myeloid leukemia - Biology & Translational Research
Keyword(s): Acute Myeloid Leukemia, FLT3, Myeloid malignancies, Myeloproliferative disorder
Abstract: PB1667
Type: Publication Only
Background
Myeloid malignancies comprise many different subtypes amongst which myeloproliferative neoplasms (MPN), myelodysplastic syndrome (MDS), myelodysplastic/myeloproliferative neoplasms (MDS/MPN) and acute myeloid leukemia (AML), are associated with somatic mutations acquired in around 20-30 key genes. To detect these relevant myeloid malignancy-related genetic variants, a targeted enrichment approach coupled to NGS analysis nicely complement the individual biomarker tests to provide a comprehensive mutational overview.
The QIAact Myeloid DNA UMI Panel in combination with the QIAGEN GeneReader™ NGS System provides an integrated solution to simultaneously interrogate many candidate genes for actionable mutations, from as little as 40 ng input DNA, shortening test time and enabling simplification of lab operations.
The QIAact Myeloid DNA UMI Panel is a 25-gene targeted sequencing panel for markers of known significance to clonal myeloid malignancies, allowing reliable and sensitive detection of single nucleotide variants (SNV) and large Insertion/Deletion (InDel) mutations.
Aims
To assess the QIAact Myeloid DNA UMI Panel performance in combination with the QIAGEN GeneReader NGS System.
Methods
Following the advice from key opinion leaders in the field of blood cancer disorders, the QIAact Myeloid DNA UMI Panel has been designed to detect relevant mutations throughout the most informative genes linked to myeloid disease. A key feature of the panel is the addition of a unique molecular index (UMI) to tag individual molecules prior to target enrichment by PCR. This enables sequencing and PCR bias correction, allowing sensitive detection of mutations (e.g. below 1% MAF for JAK2 (exon 12, 13, 14 & 15) & KIT (exon 8, 9, 10, 11 & 17)).
To assess the assay performance, control samples and blood and bone marrow samples were used. Following target enrichment, libraries were sequenced on the GeneReader™ NGS system and mutations identified using the QIAGEN Clinical Insight (QCI™) Analyze software suite, adjusted to specifically support variant calling in this assay.
Results
Complex mutations like the 52 bp deletion CALR type 1 variant, FLT3 ITDs up to 121 bp insertion and NPM1 insertions were efficiently identified. Limit of detection testing using the power of UMIs demonstrated uniform amplification and sequencing coverage to consistently detect mutations with a 1% MAF and below for JAK2 and KIT (KIT D816V variant can be detected at 0.2%), and 5% for all the other genes covered by the panel. Overall mutational variants were correctly called, resulting in >99% concordance with an alternative testing technology.
Conclusion
The QIAact Myeloid DNA UMI Panel in combination with the QIAGEN GeneReader NGS System offer a fully integrated sample to insight solution. The optimized chemistry allows superior analytical sensitivity in accurately detecting highly relevant genetic alterations for myeloid malignancy research.
Session topic: 3. Acute myeloid leukemia - Biology & Translational Research
Keyword(s): Acute Myeloid Leukemia, FLT3, Myeloid malignancies, Myeloproliferative disorder