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Updated: Jun 7, 2026
Multiplexed Fluorescent Immunohistochemical Staining, Imaging, and Analysis in Histological Samples of Lymphoma
Published on: January 9, 2019
Detection of clonality in follicular lymphoma using formalin-fixed, paraffin-embedded tissue samples and BIOMED-2
Ellen Berget1, Lars Helgeland, Anders Molven
1Department of Pathology, The Gade Institute, Haukeland University Hospital, Bergen, Norway.
Aims:
The BIOMED-2 multiplex PCR protocol is a commonly used procedure for assessing B cell clonality in lymphoma diagnostics. Follicular lymphoma poses a special challenge for PCR-based analyses because of high prevalence of somatic hypermutations in the rearranged immunoglobulin (IG) domains. This study aimed to evaluate the BIOMED-2 protocol performance in detection of B cell clonality in follicular lymphoma using formalin-fixed, paraffin-embedded (FFPE) tissue.
Methods:
FFPE samples from 118 patients diagnosed with follicular lymphoma in the period 1998-2008 were used in the study. Clonality of IG heavy (IGH) and light chains (IGK, IGL) was assessed using a PCR procedure that was optimised for FFPE tissue.
Results:
The highest clonal detection rates were 67.8% with the IGH Vн-FR2-Jн assay and 66.1% with the IGK Vκ-Jκ assay. Clonality was detected in 94.9% of all FFPE follicular lymphoma samples when all assays were combined. FFPE samples stored for 1-5 years did not perform significantly differently from those stored for 6-11 years. Interobserver agreement of clonality was tested for all analyses. The lowest score (Cohen's κ value = 0.56) was observed for the IGK Vκ-Jκ clonality assay.
Conclusions:
An improved PCR protocol for detection of clonality in FFPE samples using BIOMED-2 IG primers is presented. For best performance, a combination of IGH and IGK analyses is recommended.
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