The Ion Torrent Oncomine BCR IGH LR Assay is a robust, targeted next-generation sequencing (NGS) assay designed for use in hematology-oncology, immuno-oncology, infectious disease, and basic immunology research. Unlike other technologies, the Oncomine BCR IGH-LR Assay offers more than 400 base amplicons of library sequencing through long-read sequencing chemistry, enabling comprehensive coverage of the B-cell receptor (BCR) immunoglobin heavy (IGH) chain. The assay kit includes a single pool of multiplex PCR primers, cDNA synthesis kit, and Ion Ampliseq library reagents.
The Oncomine BCR IGH LR Assay is designed to accurately measure clonal expansion, quantify somatic hypermutation, and isotype B lymphocytes using total RNA extracted from bone marrow, whole blood, peripheral blood leukocytes (PBLs), peripheral blood mononuclear cells (PBMCs), or sorted cells in fresh-frozen research specimens. The assay utilizes Ion AmpliSeq multiplex PCR technology. It amplifies framework 1 (FR1) and the constant gene region of the BCR IGH gene to interrogate highly diverse complementarity-determining regions CDR1, CDR2, and CDR3, as well as the CH1 domain of the constant gene to enable distinction of all nine isotypes. IGH CDR-region amino acid motifs may reveal signatures of B-cell responses to defined antigens for use as future markers of autoimmune or infectious disease. Automated clonal lineage analysis and multi-sample analysis capability facilitates tracking of B-cell clonal evolution arising from somatic hypermutation and class switch recombination in research samples.
Oncomine BCR IGH LR Assay benefits include: • Extended amplicon allows for quantification of variable gene somatic hypermutation and isotype identification to enable hemato-oncology, immuno-oncology, and infectious disease translational and clinical research • Assay primers are designed to amplify all variable and joining gene alleles in the gold-standard IMGT database • RNA input improves detection of changes in plasmablast and plasma cell populations in research samples following immune challenge or administration of immunomodulatory agents • Superior multiplex PCR design through Ion AmpliSeq technology assures high accuracy and high sensitivity • Ultra-low substitution error rate minimizes artifacts arising from sequencing errors to enable highly accurate assessment of somatic hypermutation and clonal heterogeneity in malignant B-cell research samples of interest • The entire workflow from isolation of RNA to analysis of samples can be accomplished within two days using the Ion Chef templating system and Ion Genestudio S5 sequencing system • Flexible input requirements ensure successful library construction with as low as 25 ng and up to 2 µg total RNA input • Streamlined and user-friendly informatics solution offers automated clonotyping, somatic hypermutation quantification, clonal lineage analysis, reporting of key repertoire features, and multi-sample analysis capability to track B-cell clones across research samples