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Molecular pathology diagnosis of diffuse large B cell lymphoma using BIOMED-2 clonal gene rearrangements
1Department of Molecular Biology, Ahar Branch, Islamic Azad University, Ahar, Iran.
Insights
Clonality testing using BIOMED-2 protocols on FFPE tissues effectively detects immunoglobulin gene rearrangements in diffuse large B cell lymphoma (DLBCL). This molecular analysis significantly improves DLBCL diagnosis and aids in identifying lymphoproliferative disorders.
Area of Science:
- Oncology
- Molecular Biology
- Immunology
Background:
- Clonality testing for immunoglobulin gene rearrangement analysis is a valuable diagnostic technique for lymphoma.
- The BIOMED-2 project established a gold standard method for clonality detection.
Purpose of the Study:
- To empirically test clonality rearrangements of IGH and incomplete IGH D-J on formalin-fixed, paraffin-embedded (FFPE) tissues from diffuse large B cell lymphoma (DLBCL) patients.
- To evaluate the effectiveness of BIOMED-2 protocols for clonality detection in DLBCL.
Main Methods:
- Analysis of 50 sequential FFPE specimens from DLBCL patients.
- Standard multiplex PCR and heteroduplex techniques were used to analyze IGH and incomplete IGH D-J clonal gene rearrangements.
Main Results:
- A high rate of positive monoclonality was identified: 96% for IGH and 58% for incomplete IGH D-J.
- Specific rates for IGH frameworks (FRIII, FRII, FRI) and incomplete IGH D-J (DH1-6-JH, DH7-JH) were determined.
- No gene rearrangements were detected in 4% of cases.
Conclusions:
- Molecular clonality gene rearrangement analysis on FFPE tissues using BIOMED-2 protocols significantly improves clinicopathological diagnosis of DLBCL.
- Clonality testing is a reliable technique for routine diagnosis of DLBCL and other lymphoproliferative disorders.
Background:
Clonality testing for immunoglobulin gene rearrangement analysis could be implemented effectively as a useful technique for conventional diagnosis of lymphoma. The European Biomedicine and Health Concerted Action Project BMH4-CT98-3936 (BIOMED-2) have been suggested a gold standard method to clonality detection.
Objectives:
We tested empirically clonality rearrangements of IGH and incomplete IGH D-J, on formalin-fixed, paraffin embedded (FFPE) tissue of patients with diffuse large B cell lymphoma (DLBCL).
Material And Methods:
This appraisal was conveyed on 50 sequential FFPE specimens of patients with DLBCL. We carried out a standard multiplex PCR and heteroduplex techniques to analysis of IGH and incomplete IGH D-J clonal gene rearrangements.
Results:
In our investigation, we were identified a total positive monoclonality of 96% (48/50) for IGH and 58% (29/50) for incomplete IGH D-J. The percentage of positive clonality was detected in three frameworks (FRI, II, III) of IGH revealed 50% (25/50), 28% (14/50) and 18% (9/50) to FRIII, FRII and FRI, respectively. Analysis of incomplete IGH D-J showed 34% (17/50) and 24% (12/50) rates of positive clonality for DH1-6-JH and DH7-JH, respectively. In the 4% (2/50) of cases was no detected any gene rearrangements in both of IGH and incomplete IGH D-J genes.
Conclusions:
Analysis of molecular clonality gene rearrangements on FFPE tissues disclosed that using BIOMED-2 protocols, could be improvement significant clinicopathological diagnosis of DLBCL. Clonality testing is believable that to suggest as a helpful and credible technique for clonality detection in the routine diagnosis of DLBCL and other lymphoproliferative disorders.
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