A practical strategy for the routine use of BIOMED-2 PCR assays for detection of B- and T-cell clonality in diagnostic haematopathology

Br J Haematol. 2007 Jul;138(1):31-43. doi: 10.1111/j.1365-2141.2007.06618.x.

Abstract

BIOMED-2 polymerase chain reaction (PCR) assays for clonality analysis of immunoglobulin (IG) and T-cell receptor (TCR) gene rearrangements were evaluated in routine haematopathological practice where paraffin-embedded tissues constitute the majority of specimens. One hundred and twenty-five fresh/frozen and 316 paraffin specimens were analysed for DNA quality and clonality. Seventy-nine per cent of paraffin specimens yielded PCR products of over 300 bp. These specimens and all fresh/frozen specimens were analysed with the complete set of BIOMED-2 reactions for IG (8 reactions) and/or TCR (6 reactions) gene rearrangements. The rate of detection of clonality was 96% in mature B-cell neoplasms and 98% in mature T-cell neoplasms and there were no significant differences in these rates between paraffin and fresh/frozen specimens. As the value of sole use of any individual BIOMED-2 reaction in clonality detection was limited, we assessed combinations of reactions that gave the greatest sensitivity with fewest reactions and were applicable for both fresh/frozen and paraffin specimens. For IG gene rearrangements, three reactions combining one targeting the IG heavy chain framework-2 region and two targeting the IG kappa locus achieved a 91% detection rate. For TCR gene rearrangements, the two TCR gamma reactions gave a 94% detection rate. We therefore recommend this strategy as the first-line assays for routine B- and T-cell clonality analysis in diagnostic haematopathology.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • B-Lymphocytes / pathology*
  • Clone Cells
  • DNA Primers / genetics
  • Electronic Data Processing*
  • Gene Rearrangement*
  • Hematologic Neoplasms / diagnosis*
  • Hematologic Neoplasms / genetics
  • Humans
  • Paraffin Embedding
  • Polymerase Chain Reaction / methods
  • Sensitivity and Specificity
  • T-Lymphocytes / pathology*

Substances

  • DNA Primers