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CAT #: 12060020

TCRD Gene Clonality Assay MegaKit for Gel Detection

Assay Use

This Research Use Only assay identifies T-cell receptor delta chain clonality and is useful for the study of identifying 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.1  This PCR assay employs multiple consensus DNA primers that target conserved genetic regions within the T-cell receptor delta chain gene.  The human T-cell receptor delta (TCRD) locus comprises a cluster of 10 genes located on chromosome 14 at 14q11.2.  The cluster spans 60 kb and is localized between the T-cell receptor alpha (TCRA) variable and joining region genes.

    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. Invivoscribe’s gene clonality 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.

    SUMMARY
    This test kit includes two (2) master mixes.  The TCRD Tube multiplex master mix contains 12 individual primers 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).  This standardized multiplex PCR assay detects the vast majority of clonal TCR delta gene rearrangements using a single multiplex master mix (Figure).  The Specimen Control Size Ladder master mix, targets multiple genes and generates a series of amplicons of approximately 100, 200, 300, 400, and 600 base pairs (bp) to ensure that the quality and quantity of input DNA is adequate to yield a valid result.  A single thermal cycler program and similar detection methodologies are used with all of our gene clonality assays which improves consistency and facilitates cross training on a broad range of different assays.

  • Specimen Requirements

    This assay tests genomic DNA.

References

1.  Miller, JE, et al.  Molecular Diagnostics. 1999, 4(2):101-117.

Disclaimer

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

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