Design and standardization of PCR primers and protocols for detection of clonal immunoglobulin and T-cell receptor gene recombinations in suspect ... View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2003-12

AUTHORS

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

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. More... »

PAGES

2257

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  • Journal

    TITLE

    Leukemia

    ISSUE

    12

    VOLUME

    17

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/sj.leu.2403202

    DOI

    http://dx.doi.org/10.1038/sj.leu.2403202

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1027716225

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/14671650


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