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Immunoglobulin Gene Sequence Analysis In Chronic Lymphocytic Leukemia: From Patient Material To Sequence Interpretation
Published on: November 26, 2018
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Next-Generation Sequencing-Based Clonality Assessment of Ig Gene Rearrangements: A Multicenter Validation Study by
Michiel van den Brand1, Jos Rijntjes1, Markus Möbs2
1Department of Pathology, Radboud University Medical Center, Nijmegen, the Netherlands.
The Journal of Molecular Diagnostics : JMD
|June 29, 2021
Summary
Next-generation sequencing (NGS) for immunoglobulin (IG) gene clonality analysis offers a sensitive and reliable alternative to conventional methods. This validated NGS approach shows high concordance, supporting its use in diagnosing B-cell lymphoid proliferations.
Area of Science:
- Hematology
- Molecular Biology
- Oncology
Background:
- Immunoglobulin (IG) gene clonality analysis is crucial for differentiating benign from malignant B-cell proliferations.
- Conventional methods rely on EuroClonality/BIOMED-2 multiplex PCR and GeneScan analysis.
- Next-generation sequencing (NGS) offers a novel approach to IG clonality assessment.
Purpose of the Study:
- To biologically validate a new NGS-based method for IG clonality analysis.
- To compare the performance of NGS with the established EuroClonality/BIOMED-2 protocol.
- To assess the interlaboratory concordance and diagnostic accuracy of NGS-based clonality analysis.
Main Methods:
- International multicenter validation of NGS-based IG clonality analysis.
- Comparison of 209 specimens (reactive and neoplastic lymphoproliferations) using both NGS and conventional EuroClonality/BIOMED-2 protocols.
- Analysis of immunoglobulin heavy chain and kappa light chain targets.
Main Results:
- High interlaboratory concordance (99%) and concordance with conventional analysis (98%) for NGS.
- NGS demonstrated higher sensitivity in detecting clonal rearrangements and providing interpretable results.
- Clonal rearrangements were detected in 96% of B-cell neoplasms by NGS versus 95% by conventional methods; reactive cases were predominantly polyclonal.
Conclusions:
- NGS-based IG clonality analysis is comparable in performance to conventional methods.
- The study provides parameters for interpreting NGS clonality results and suggests a path toward quantitative scoring.
- High sensitivity and clonal comparison capabilities support the diagnostic implementation of NGS-based clonality analysis.
