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.

J.J. van Dongen, A.W. Langerak, M. Brüggemann, P.A. Evans, M. Hummel, F.L. Lavender, E. Delabesse, F. Davi, E. Schuuring, R. García-Sanz, J.H. van Krieken, J. Droese, D. González, C. Bastard, H.E. White, M. Spaargaren, M. González, A. Parreira, G.J. Morgan, M. KnebaE.A. Macintyre, J.L Smith

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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.
Original languageEnglish
Pages (from-to)2257-2317
Number of pages61
JournalLeukemia
Volume17
Issue number12
DOIs
Publication statusPublished - Dec 2003

Keywords

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

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