A Novel nested-PCR Strategy for the Detection of Rearranged Immunoglobulin Heavy-Chain Genes in B Cell Tumors

Leukemia. 1997 Oct;11(10):1793-8. doi: 10.1038/sj.leu.2400801.

Abstract

Several methods have been developed for the detection of minimal residual disease (MRD) in B cell tumors. Chromosomal translocations or the rearrangement of the immunoglobulin heavy chain (IgH) and T cell receptor genes are generally employed. We report a novel PCR method to detect MRD using IgH genes. IgH rearranged variable region (VDJ) were amplified from tumor specimens using consensus primers for variable and joining region genes. Complementarity-determining regions (CDR) were identified and used to generate tumor-specific primers. Two-round amplifications using primers derived from CDRs and joining or constant regions were performed for MRD detection. IgH nested-PCR approach was tested on a panel of 75 B cell tumors including acute lymphoblastic and chronic lymphocytic leukemias, non-Hodgkin's lymphomas and multiple myelomas. A VDJ sequence was obtained in 62 out of 75 cases (83%). Sensitivity using DNA or cDNA templates was 10(-5) and (-6), respectively. This method is specific and sensitive and provides a simple, non-radioactive approach for the evaluation of MRD in B cell tumors.

Publication types

  • Research Support, Non-U.S. Gov't
  • Technical Report

MeSH terms

  • Burkitt Lymphoma / genetics*
  • DNA Primers
  • DNA, Neoplasm / analysis
  • DNA, Neoplasm / genetics
  • Gene Amplification
  • Gene Rearrangement, B-Lymphocyte, Heavy Chain*
  • Genes, Immunoglobulin*
  • Humans
  • Leukemia, Lymphocytic, Chronic, B-Cell / genetics*
  • Lymphoma, B-Cell / genetics*
  • Multiple Myeloma / genetics*
  • Neoplasm, Residual
  • Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity

Substances

  • DNA Primers
  • DNA, Neoplasm