Improved clonality detection in Hodgkin lymphoma using a semi-nested modification of the BIOMED-2 PCR assay for IGH and IGK rearrangements: A paraffin-embedded tissue study

Pathol Int. 2018 May;68(5):287-293. doi: 10.1111/pin.12660. Epub 2018 Mar 25.

Abstract

The BIOMED-2 PCR protocols targeting IGH and IGK genes may be useful for detecting clonality in Hodgkin lymphoma (HL). The clonality detection rates, however, have not been very high with these methods using paraffin-embedded tumor sections. We previously described the usefulness of the semi-nested BIOMED-2 IGH assay in B-cell malignancies. In this study, we devised a novel semi-nested BIOMED-2 IGK assay. Employing 58 cases of classical HL, we carried out the standard BIOMED-2, BIOMED-2 followed by BIOMED-2 re-amplification, and BIOMED-2 followed by semi-nested BIOMED-2, all targeting IGH and IGK, using paraffin-embedded tissues. In both IGH and IGK assays, semi-nested assays yielded significantly higher clonality detection rates than the standard assays and re-amplification assays. Clonality was detected in 13/58 (22.4%) classical HL cases using the standard IGH/IGK assays while it was detected in 38/58 (65.5%) cases using semi-nested IGH/IGK assays. The detection rates were not associated with the HL subtypes, CD30-positive cell density, CD20-positive cell density, or Epstein-Barr virus (EBV) positivity. In conclusion, tumor clonality was detected in nearly two-thirds of classical HL cases using semi-nested BIOMED-2 IGH/IGK assays using paraffin tumor sections. These semi-nested assays may be useful when the standard IGH/IGK assays fail to detect clonality in histopathologically suspected HLs.

Keywords: BIOMED-2 PCR assay; Hodgkin lymphoma; IGH gene; IGK gene; formalin-fixed; immunoglobulin gene rearrangements; paraffin-embedded tissues; semi-nested modification.

MeSH terms

  • Clone Cells
  • Gene Rearrangement, B-Lymphocyte, Heavy Chain / genetics*
  • Gene Rearrangement, B-Lymphocyte, Light Chain / genetics*
  • Genes, Immunoglobulin Heavy Chain / genetics
  • Hodgkin Disease / genetics
  • Hodgkin Disease / immunology*
  • Humans
  • Immunoglobulin kappa-Chains / genetics
  • Paraffin Embedding
  • Reverse Transcriptase Polymerase Chain Reaction / methods*

Substances

  • Immunoglobulin kappa-Chains