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IgVH Somatic Hypermutation by Next Generation Sequencing

IgVH Somatic Hypermutation by Next Generation Sequencing

NGS Testing for IgVH Somatic Hypermutation is Superior to Traditional Methodologies:

Traditionally, IgVH SHM has been evaluated by rt-PCR followed by Sanger Sequencing using patient sample RNA. However, RNA lability places a significant burden on the submitting physician to minimize specimen transit time. The Sanger Sequencing approach is also time and labor intensive and may show limited sensitivity in detection of IgVH SHM for low abundance clones. Flow cytometric analysis of ZAP-70 expression has therefore been widely utilized as a surrogate for IgVH SHM status, however, standardization for this marker is known to be poor due to the subjective nature of its interpretation. Significant discordance between ZAP-70 expression patterns and IgVH SHM results may also be seen.

Next generation sequencing offers significant advantages in the evaluation of IgVH somatic hypermutation status in CLL/SLL
  • Utilizes patient DNA instead of RNA as sample starting material, eliminating many of the challenges related to RNA-based testing.
  • Minimal sample required: 1ml blood or bone marrow aspirate, or 0.01 µg DNA
  • Non-subjective data output includes percentage homology to germline reference IGH sequences, clonal IGH sequence abundance, and VDJ gene utilization
  • Demonstrates improved sensitivity in detection of clonal populations
Additional Resources

IgVH Information Sheet

Example IgVH SHM Report

Association for Molecular Pathology 2016 Poster: NGS demonstrates Clinical Utility and Increased Sensitivity in Detection of IgVH Somatic Hypermutation in CLL/SLL

International CLL-IPI working group.  An international prognostic index for patients with chronic lymphocytic leukaemia (CLL-IPI): a meta-analysis of individual patient data.  Lancet Oncol. 2016 Jun;17(6):779-90.

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