title: 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.
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.
authors.affiliation: Department of Immunology, Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands. j.j.m.vandongen@erasmusmc.nl
authors.forename: J J M
authors.initials: JJ
authors.lastname: van Dongen
authors.forename: A W
authors.initials: AW
authors.lastname: Langerak
authors.forename: M
authors.initials: M
authors.lastname: Brüggemann
authors.forename: P A S
authors.initials: PA
authors.lastname: Evans
authors.forename: M
authors.initials: M
authors.lastname: Hummel
authors.forename: F L
authors.initials: FL
authors.lastname: Lavender
authors.forename: E
authors.initials: E
authors.lastname: Delabesse
authors.forename: F
authors.initials: F
authors.lastname: Davi
authors.forename: E
authors.initials: E
authors.lastname: Schuuring
authors.forename: R
authors.initials: R
authors.lastname: García-Sanz
authors.forename: J H J M
authors.initials: JH
authors.lastname: van Krieken
authors.forename: J
authors.initials: J
authors.lastname: Droese
authors.forename: D
authors.initials: D
authors.lastname: González
authors.forename: C
authors.initials: C
authors.lastname: Bastard
authors.forename: H E
authors.initials: HE
authors.lastname: White
authors.forename: M
authors.initials: M
authors.lastname: Spaargaren
authors.forename: M
authors.initials: M
authors.lastname: González
authors.forename: A
authors.initials: A
authors.lastname: Parreira
authors.forename: J L
authors.initials: JL
authors.lastname: Smith
authors.forename: G J
authors.initials: GJ
authors.lastname: Morgan
authors.forename: M
authors.initials: M
authors.lastname: Kneba
authors.forename: E A
authors.initials: EA
authors.lastname: Macintyre
mesh_terms: D002869:Chromosome Aberrations; D002999:Clone Cells; D017931:DNA Primers; D005062:European Union; D015329:Gene Rearrangement, T-Lymphocyte; D006801:Humans; D007136:Immunoglobulins / Q000235:genetics*; D008232:Lymphoproliferative Disorders / Q000175:diagnosis* / Q000235:genetics*; D018365:Neoplasm, Residual / Q000175:diagnosis / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards*; D011948:Receptors, Antigen, T-Cell / Q000235:genetics*; D012015:Reference Standards; D015203:Reproducibility of Results
chemical_list: D017931:DNA Primers; D007136:Immunoglobulins; D011948:Receptors, Antigen, T-Cell