Design and Standardization of PCR Primers and Protocols for Detection of Clonal Immunoglobulin and T-cell Receptor Gene Recombinations in Suspect Lymphoproliferations: Report of the BIOMED-2 Concerted Action BMH4-CT98-3936

Leukemia. 2003 Dec;17(12):2257-317. doi: 10.1038/sj.leu.2403202.

Abstract

In a European BIOMED-2 collaborative study, multiplex PCR assays have successfully been developed and standardized for the detection of clonally rearranged immunoglobulin (Ig) and T-cell receptor (TCR) genes and the chromosome aberrations t(11;14) and t(14;18). This has resulted in 107 different primers in only 18 multiplex PCR tubes: three VH-JH, two DH-JH, two Ig kappa (IGK), one Ig lambda (IGL), three TCR beta (TCRB), two TCR gamma (TCRG), one TCR delta (TCRD), three BCL1-Ig heavy chain (IGH), and one BCL2-IGH. The PCR products of Ig/TCR genes can be analyzed for clonality assessment by heteroduplex analysis or GeneScanning. The detection rate of clonal rearrangements using the BIOMED-2 primer sets is unprecedentedly high. This is mainly based on the complementarity of the various BIOMED-2 tubes. In particular, combined application of IGH (VH-JH and DH-JH) and IGK tubes can detect virtually all clonal B-cell proliferations, even in B-cell malignancies with high levels of somatic mutations. The contribution of IGL gene rearrangements seems limited. Combined usage of the TCRB and TCRG tubes detects virtually all clonal T-cell populations, whereas the TCRD tube has added value in case of TCRgammadelta(+) T-cell proliferations. The BIOMED-2 multiplex tubes can now be used for diagnostic clonality studies as well as for the identification of PCR targets suitable for the detection of minimal residual disease.

Publication types

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

MeSH terms

  • Chromosome Aberrations
  • Clone Cells
  • DNA Primers
  • European Union
  • Gene Rearrangement, T-Lymphocyte
  • Humans
  • Immunoglobulins / genetics*
  • Lymphoproliferative Disorders / diagnosis*
  • Lymphoproliferative Disorders / genetics*
  • Neoplasm, Residual / diagnosis
  • Neoplasm, Residual / genetics
  • Polymerase Chain Reaction / methods*
  • Polymerase Chain Reaction / standards*
  • Receptors, Antigen, T-Cell / genetics*
  • Reference Standards
  • Reproducibility of Results

Substances

  • DNA Primers
  • Immunoglobulins
  • Receptors, Antigen, T-Cell