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CAT #: 12070111

T Cell Receptor Gamma Gene Rearrangement Assay 2.0 MegaKit

Assay Use

This Research Use Only assay identifies T-cell receptor gamma (TCRG) chain gene rearrangements and is useful for the identification of clonal T-cell populations and evaluation of new research and methods in malignancy studies.

Product Details

  • Summary and Explanation of the Test

    BACKGROUND
    Invivoscribe’s T-Cell Receptor Gamma Gene Rearrangement Assay 2.0 represents an improved approach to PCR-based clonality testing of T-cell lymphoproliferative disorders.  This assay was optimized using a single amplification reaction with a single fluorescent dye for detection. Amplified products generated targeting the TCR gamma gene locus all fall within a single size range to facilitate interpretation.

    The human TCR gamma gene locus on chromosome seven (7q14) includes 14 V genes (six of these V genes are functional; three (3) open reading frames and five (5) pseudogenes) belonging to four subgroups (Group I, II, III and IV), five (5) J segments and two (2) C genes spread over 200 kilobases.  The diversity of this locus has complicated PCR-based testing.  This multiplex PCR assay represents an improvement over existing assays as it can detect the vast majority of TCR gamma gene rearrangements with a single multiplex master mix.  Importantly, this assay includes primers for all known groups of TCR gamma variable region genes and joining region genes involved in rearrangements in T-cell lymphomas.  In addition, all labeled primers are conjugated with the 6FAM fluorophore.

    SUMMARY
    Our single multiplex master mix targets all conserved regions within the variable (V) and the joining (J) region genes that are described in lymphoid malignancies.  This is critical for more comprehensive analysis of samples, as some T-cell lymphoproliferative disorders involve V and J segments that would not be identified with a single Vγ (1-8) and Jγ1/2 primer set. Amplification with all primers in a single tube has several additional important advantages over existing methods.  The polyclonal background that results from the combination of all primers in a single tube produces a more robust and easily interpreted signal with capillary electrophoresis, which aids in the interpretation of small peaks. The average size of the TCRG gene rearrangement PCR product is 190 bp, with a normal distribution of product sizes between 159 and 207 bp.  This protocol leads to improved PCR product formation from paraffin-embedded samples compared to other protocols that yield products of 260 bp or longer.  Positive and negative DNA controls, as well as a Specimen Control Size Ladder master mix are included.  Clonality is indicated if a dominant amplicon is detected.

    This test kit includes the TCRG – 6FAM master mix that targets framework regions within the variable region and the joining region of the TCR gamma chain locus.  The second master mix, the Specimen Control Size Ladder, targets multiple genes and generates a series of amplicons of 100, 200, 300, 400, and 600 bp that can be used to ensure that the quality 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; OR
    • Formalin-fixed paraffin embedded tissue or slides.

Disclaimer

This assay was developed by Invivoscribe.  The performance of this assay was reviewed and validated by the EuroClonality/BIOMED-2 Group.

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