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CAT #: 12070051

T-Cell Receptor Gamma Gene Rearrangement Assay for ABI Fluorescence Detection

Assay Use

This Research Use Only assay identifies T-Cell Receptor Gamma 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
    Rearrangement of the variable (Vγ) and joining (Jγ) gene segments of the TRG locus results in Vγ-Jγ products of unique length and sequence.  Clonal TRG rearrangements can be most rapidly identified by analyzing the size distribution of DNA products amplified from conserved sequences that flank this Vγ-Jγ region.1  DNA isolated from a normal heterogeneous population of polyclonal T-cells produces a Gaussian distribution (bell-shaped size curve) of amplified products.  DNA amplified from a clonal T-cell population generates one or two product(s) of unique size that reflects proliferation of a single rearranged clone.1,2

    The T-cell receptor gamma (TRG, formerly known as TCRG) chain locus spans 160 kilobases on chromosome 7 (7p14).  The locus consists of 14 Vγ gene segments in six (6) subgroups, and five (5) Jγ gene segments interspersed between two (2) constant (Cγ) gene segments.  However, the repertoire of functional TRG molecules is limited to 4-6 functional Vγ gene segments that belong to two (2) subgroups.2

    SUMMARY
    Invivoscribe’s gene rearrangement and translocation assays represent a simple approach to PCR-based testing and utilize a series of standardized master mixes that have been carefully optimized.  Several master mixes are used to test for rearrangements at each receptor locus and each master mix targets a different conserved region within the receptor gene.  This comprehensive testing approach maximizes identification of clonal rearrangements.  A single universal thermal cycler program and similar detection methodologies are used with all of our tests; this improves consistency and facilitates cross-training.

    This test kit includes three (3) master mixes. TCRG Mix 1 and TCRG Mix 2 target framework regions within the variable (V) region and the joining (J) region of the TCR gamma chain locus.  The Amplification Control master mix targets the HLA-DQα locus and generates an amplicon of 235 base pairs (bp) to ensure the quality and quantity of input DNA is adequate to yield a valid result.

  • Specimen Requirements

    This assay tests extracted and purified genomic DNA (gDNA).  Common sources of gDNA include:

    • 5 cc of peripheral blood, bone marrow biopsy or bone marrow aspirate anti-coagulated with heparin or EDTA. Ship at ambient temperature; OR
    • Formalin-fixed paraffin embedded tissue or slides.

References

1. Miller, JE et al. (1999) Molecular Diagnostics. 4(2):101-117.
2. van Dongen, JJM et al. (2003) Leukemia. 17(12):2257-2317.

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