Detection of clonal B cell populations in paraffin-embedded tissues by polymerase chain reaction

Am J Pathol. 1993 Jul;143(1):230-9.

Abstract

A method was established to detect clonal B cell populations in frozen and paraffin-embedded tissues. The method is based on the polymerase chain reaction amplification of rearranged VH and V kappa genes using V gene family-specific primers. Monoclonal B cell populations could be detected in peripheral blood lymphocyte DNA in all 16 cases of B-CLL and immunocytoma investigated and in 8 of 10 cases of B cell non-Hodgkin's lymphoma using frozen and paraffin sections. The amplification of V kappa rearrangements in addition to the VH amplification is a useful tool to verify the results of the heavy chain rearrangement and to detect proliferation of a B cell clone in cases in which no VH product was obtained. In spite of the degradation of DNA in paraffin-embedded tissues, we were able to find amplified polymerase chain reaction products of about 350 bp length in 8 of 10 cases analyzed. The method presented here may be helpful in routine diagnosis of B cell non-Hodgkin's lymphoma using frozen or paraffin-embedded specimens.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • B-Lymphocyte Subsets / cytology*
  • Base Sequence
  • Blotting, Southern
  • Clone Cells / cytology*
  • Electrophoresis, Agar Gel
  • Frozen Sections
  • Gene Amplification
  • Gene Rearrangement, B-Lymphocyte, Heavy Chain
  • Gene Rearrangement, B-Lymphocyte, Light Chain
  • Humans
  • Immunoglobulin Heavy Chains / genetics
  • Immunoglobulin Light Chains / genetics
  • Leukemia, B-Cell / genetics
  • Molecular Sequence Data
  • Paraffin Embedding
  • Polymerase Chain Reaction

Substances

  • Immunoglobulin Heavy Chains
  • Immunoglobulin Light Chains