Race Against the Clock: Validating a Rapid NGS Myeloid Panel
The Hospital of the University of Pennsylvania built its Hematological Expedited Sequencing (HEXS) assay on Pillar’s oncoReveal® Myeloid Panel, consolidating 58 myeloid neoplasm genes into a single NGS test to replace multiple send-outs for acute leukemia subtyping. Validation used 61 positive and 18 negative bone marrow, peripheral blood, and bone core specimens covering SNVs, indels, and FLT3 internal tandem duplications, on both the MiSeq and MiSeq i100 with PiVAT® variant calling. HEXS reached 99.5–100% sensitivity across variant types and enables 3–5 day myeloid NGS reporting, with the i100 cutting sequencing time by 17 hours.
Performance Comparison of the Illumina MiSeq i100 with Established Instruments for Amplicon-Based Somatic and Liquid Biopsy Sequencing Panels
Pillar benchmarked seven oncoReveal® amplicon NGS panels — BRCA1/BRCA2 Somatic with CNV, Multi-Cancer v4 with CNV, Myeloid, Essential MPN, Nexus 21 Gene, Rapid AML, and Essential LBx — on Illumina’s new MiSeq i100 against the legacy MiSeq and NextSeq 550. Fifty-nine paired samples, including SeraCare and Horizon Discovery reference standards, cancer cell line DNA, and biobanked clinical specimens, were run from identical libraries across six sequencing runs. Every expected true-positive variant was detected on the i100 with no false positives, confirming that the portfolio transfers cleanly to the new platform.
Evaluation of oncoReveal Essential MPN Panel on Illumina MiSeq i100 for Key Driver Mutation Testing in Myeloproliferative Neoplasms
Mayo Clinic’s Molecular Hematopathology Laboratory evaluated Pillar’s oncoReveal® Essential MPN Panel — a single-tube SLIMamp® amplicon NGS assay targeting JAK2, CALR, and MPL — on the Illumina MiSeq i100 for BCR::ABL1-negative myeloproliferative neoplasm (MPN) testing. Ninety-four peripheral blood and bone marrow specimens were compared against the laboratory’s capture-based myeloid NGS panel, JAK2 V617F allele-specific PCR, CALR fragment analysis, and MPL Sanger sequencing. The panel showed 91–100% concordance across comparators and returned results within 24 hours of extraction, against 5–7 days for the conventional reflex cascade.