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CAT #: 12050011

TCRB Gene Clonality Assay for ABI Fluorescence Detection

Assay Use

This Research Use Only assay uses PCR to detect clonal T-cell receptor beta chain gene rearrangements and is useful for the study of clonal T-cell populations and evaluation of new research and methods in malignancy studies.

Product Details

  • Summary and Explanation of the Test

    BACKGROUND
    Rearrangements of the antigen receptor genes occur during ontogeny in B and T lymphocytes. These gene rearrangements generate products that are unique in length and sequence for each cell.  Therefore, polymerase chain reaction (PCR) assays can be used to identify lymphocyte populations derived from a single cell by detecting the unique V-J gene rearrangements present within these antigen receptor loci.  The human TCR beta gene locus on chromosome 7 (7q35) includes 64-67 V genes belonging to 32 subgroups, 2 D segments and 13 J segments, spread over 620 kilobases.  This assay employs multiple consensus DNA primers that target conserved genetic regions within the T-cell receptor beta chain gene.  This test is used to detect the vast majority of clonal T-cell malignancies from DNA.  Test products can be analyzed using a variety of detection formats, including gel and capillary electrophoresis.

    SUMMARY
    Invivoscribe’s gene clonality and translocation assays represent a simple approach to PCR-based clonality testing.  These standardized assays were carefully optimized testing positive and negative control samples using multiplex master mixes.  This assay kit includes three (3) multiplex master mixes that target conserved regions within the variable (V), diversity (D) and the joining (J) regions that flank the unique hypervariable antigen-binding region 3 (CDR3).  TCRB Tube A contains 23 Vβ primers + 6 Jβ1 primers + 3 Jβ2 primers.  TCRB Tube B contains 23 Vβ + 4 Jβ2 primers.  TCRB Tube C contains 2 Dβ + 13 Jβ primers (Figure).  Positive and negative DNA controls, as well as an internal Specimen Control Size Ladder master mix are included.  PCR products can be analyzed by differential fluorescence detection using capillary electrophoresis or gene sequencing instruments, by heteroduplex analysis or using standard gel electrophoresis with ethidium staining.  Clonality is indicated if any one of the master mixes generates clonal band(s).

  • Specimen Requirements

    This assay tests genomic DNA.

Disclaimer

This assay is based on the EuroClonality/BIOMED-2 Concerted Action BMH4-CT98-3936.

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