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Updated: Jun 2, 2026
VDJ-Seq: Deep Sequencing Analysis of Rearranged Immunoglobulin Heavy Chain Gene to Reveal Clonal Evolution Patterns of B Cell Lymphoma
Published on: December 28, 2015
[High-throughput sequencing in identifying somatic hypermutation in immunoglobulin heavy chain variable regions with
1Peking University People's Hospital, Peking University Institute of Hematology, National Clinical Research Center for Hematologic Disease, Beijing Key Laboratory of Hematopoietic Stem Cell Transplantation, 100044 Beijing, China.
Next-generation sequencing (NGS) offers superior accuracy over Sanger sequencing for assessing immunoglobulin heavy chain variable region (IGHV) gene somatic hypermutation (SHM) status, especially in complex clonal backgrounds. NGS enhances the diagnosis and prognosis of lymphoid malignancies.
Area of Science:
- Molecular Biology
- Genetics
- Oncology
Background:
- Somatic hypermutation (SHM) of immunoglobulin heavy chain variable region (IGHV) genes is crucial for diagnosing and prognosing lymphoid malignancies.
- Accurate assessment of IGHV SHM status is vital for clinical decision-making in hematologic cancers.
- Existing methods may face challenges in complex clonal scenarios.
Purpose of the Study:
- To compare the performance of next-generation sequencing (NGS) and Sanger sequencing in evaluating IGHV gene SHM.
- To identify factors contributing to discrepancies between NGS and Sanger sequencing, particularly in complex clonal backgrounds.
- To inform optimized strategies for clinical application of these sequencing methods.
Main Methods:
- Retrospective analysis of 53 patient samples (35 chronic lymphocytic leukemia, 9 diffuse large B-cell lymphoma, etc.).
- Both Sanger sequencing and NGS were employed to assess IGHV SHM status.
- Discordant cases underwent in-depth analysis, considering clonal abundance, primer design, and interpretation criteria.
Main Results:
- NGS identified biclonal or polyclonal profiles in 23 cases initially classified as non-monoclonal by Sanger sequencing.
- NGS detected biclonality or different IGHV rearrangement types in 6 cases initially classified as monoclonal by Sanger sequencing.
- Minor differences in SHM percentage were observed, but did not significantly alter overall mutational status determination.
Conclusions:
- NGS demonstrates superior performance compared to Sanger sequencing for IGHV SHM assessment, especially in complex clonal conditions.
- NGS offers enhanced sensitivity and accuracy in detecting subclonal components and quantifying clonal proportions.
- NGS provides a more precise molecular basis for diagnosing and prognosing lymphoid malignancies, including chronic lymphocytic leukemia (CLL).
