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Updated: May 13, 2026
T and B Cell Receptor Immune Repertoire Analysis using Next-generation Sequencing
Published on: January 12, 2021
Immunoglobulin/T-cell receptor clonality diagnostics
Anton W Langerak1, Patricia Jta Groenen, J Han Jm van Krieken
1Erasmus MC, University Medical Center, Department of Immunology, Dr Molewaterplein 50, 3015 GE Rotterdam, The Netherlands +31 10 40 88089 ; +31 10 40 89456 ; a.langerak@erasmusmc.nl.
Insights
Clonality testing for lymphoid malignancies is now accessible with multiplex PCR. Accurate interpretation requires expertise and integration with pathology and immunophenotyping for reliable diagnoses.
Area of Science:
- Hematology
- Molecular Biology
- Oncology
Background:
- Clonality testing for lymphoid malignancies is crucial for diagnosis and treatment.
- Standardized multiplex polymerase chain reaction (PCR) protocols for immunoglobulin (Ig) and T-cell receptor (TCR) gene analysis facilitate routine laboratory testing.
Purpose of the Study:
- To review the essential expertise and biological knowledge required for accurate interpretation of Ig/TCR clonality data.
- To identify and discuss potential immunobiologic and technical pitfalls that can lead to misinterpretation.
- To emphasize the importance of integrating molecular data with histopathology and flow cytometry for a comprehensive diagnostic approach.
Main Methods:
- Review of current literature and established protocols for Ig/TCR gene analysis.
- Discussion of common challenges and sources of error in clonality testing.
- Exploration of multidisciplinary diagnostic models.
Main Results:
- Multiplex PCR for Ig/TCR gene analysis is technically feasible in routine settings.
- Interpretation of clonality data demands specialized knowledge of immunology and molecular biology.
- Potential pitfalls in data interpretation include technical issues and biological variations.
Conclusions:
- Accurate clonality diagnostics rely on both technical proficiency and biological understanding.
- Integrating molecular findings with pathology and immunophenotyping is vital for reliable diagnoses.
- A multidisciplinary approach ensures comprehensive data integration for optimal patient care.
Abstract:
Clonality testing in lymphoid malignancies has become technically relatively easy to perform in routine laboratories using standardized multiplex polymerase chain reaction protocols for Ig/T-cell receptor (TCR) gene analysis. Expertise with clonality diagnostics and knowledge about the biology of Ig/TCR recombination are essential for correct interpretation of the Ig/TCR clonality data. Several immunobiologic and technical pitfalls that should be taken into account to avoid misinterpretation of data are addressed in this review. Furthermore, the need to integrate the molecular data with that from (hemato-)pathology, and preferably also flowcytometric immunophenotyping for appropriate interpretation, is discussed. Such an interactive, multidisciplinary diagnostic model guarantees integration of all available data to reach the most reliable diagnosis.
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