Clinical Verification of an FDA-Approved NGS Solid Tumor IVD Assay
Evaluation of the QIAxcel Connect system for NGS library prep QC analysis: experience from a clinical diagnostic laboratory
Carolinas Pathology Group and Atrium Health evaluated the QIAGEN QIAxcel Connect system for library quantification and quality control across three NGS workflows: Archer’s FUSIONPlex Pan Solid Tumor v2, Pillar’s oncoReveal® Solid Tumor v2, and Pillar’s oncoReveal® Essential MPN panel. Libraries from 160 FFPE scrolls, 134 FFPE slides, and 126 blood or bone marrow specimens were normalized on QIAxcel concentrations, pooled per panel, and sequenced on the Illumina NextSeq 550 Dx. Pillar panels showed the tightest peak-size reproducibility, and the approach removed the need to quantify every individual library.
Analytical Validation of oncoReveal CDx as a FDA approved, Pan-Cancer Solid Tumor IVD Assay
oncoReveal® CDx is the first FDA-approved distributed, kit-based NGS assay for both companion diagnostic use and pan-cancer tumor profiling, covering more than 3,600 SNV and indel targets across 22 genes from FFPE DNA on the Illumina MiSeqDx. Validation spanned 10 tumor types and addressed DNA input range, minimum tumor content, necrosis tolerance, limit of detection by probit analysis, accuracy against two externally validated NGS comparators, and a three-site reproducibility study. The assay achieved 100% positive and 99.996% negative percent agreement at 30 ng input, with valid results down to 3.3% tumor content.
Automation of oncoReveal Solid Tumor 22 Gene Panel and the Illumina TruSight Oncology 500 DNA/RNA
Beckman Coulter Life Sciences automated both Pillar’s oncoReveal® Solid Tumor 22 Gene Panel and Illumina’s TruSight Oncology 500 DNA/RNA kit on the Biomek NGeniuS library preparation system to reduce manual pipetting and hands-on time. Three test runs per assay used engineered controls including Horizon Discovery fcDNA and SeraSeq® reference standards, sequenced on Illumina MiSeq and NextSeq 500 and analyzed with PiVAT® and DRAGEN. The Pillar panel reached sequencing by the end of day one at batch sizes from 4 to 24 samples, while the 523-gene TSO500 workflow ran into day three.
JAK2 Exon 12-15, CALR and MPL Essential MPN NGS Panel
Labcorp’s Center for Molecular Biology and Pathology validated Pillar’s oncoReveal® Essential MPN Panel — a single-tube SLIMamp® assay covering JAK2 exons 12–15, CALR, and MPL — as a high-throughput clinical test for BCR/ABL-negative myeloproliferative neoplasms including polycythemia vera, essential thrombocythemia, and primary myelofibrosis. Across 126 blood, bone marrow, and cell pellet specimens every reportable mutation was confirmed by a secondary method, giving 100% concordance, with 100% repeatability and reproducibility over 30 specimens. The panel is now offered clinically, and average laboratory turnaround shortened by four days versus cascading reflex orders.