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CAT #: 12070101

T Cell Receptor Gamma Gene Rearrangement Assay 2.0 - ABI Fluorescence Detection

Product Use

T Cell Receptor Gamma Chain Gene Rearrangement Assays are useful for the study of:

  • Identifying clonal T-cell populations highly suggestive of T-cell malignancies
  • Monitoring and evaluation of disease recurrence
  • Evaluation of new research and methods in malignancy studies

Product Details

  • Summary of Explanation of the Test

    SUMMARY:

    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.

    BACKGROUND:

    The human TCR gamma gene locus on chromosome 7 (7q14) includes 14 V genes (6 of these V genes are functional; 3 open reading frames, and 5 pseudogenes) belonging to 4 subgroups (Group I, II, III, and IV), 5 J segments, and 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. The assay provides rapid TCRG clonality assessment in 4-6 hours.

    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 patient samples, as some T-cell lymphoproliferative disorders involve V and J segments that would not be identified with a single Vg (1-8) and Jg1/2 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.  Competitive amplification of all TCRG gene rearrangements allows for identification of a quantitative threshold for a positive result and helps to avoid false positive results.  The average size of the TCRG gene rearrangement PCR amplicons is 190 nucleotides, with a normal distribution of product sizes between 159 and 207 nucleotides.  This protocol should lead to improved product formation from paraffin-embedded samples compared to other protocols that yield products of 260 nucleotides 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.

    Capillary electrophoresis provides a powerful method for the detection of amplicons that have been labeled with primers conjugated with fluorescent dyes. Reaction products are fractionated and detected simultaneously using the capillary electrophoresis. This detection system results in unsurpassed sensitivity and product resolution.

    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 base pairs to ensure that the quality of input DNA is adequate to yield a valid result. This robust Invivoscribe assay can be used to test DNA extracted from virtually any source.

  • Specimen Requirements

    This assay tests genomic DNA

    1. 5 cc of peripheral blood, bone marrow biopsy, or bone marrow aspirate anti-coagulated with heparin or EDTA
    2. Minimum 5 mm cube of tissue
    3. 100 ng of genomic DNA
    4. Formalin-fixed paraffin embedded tissue or slides

Disclaimer

This product is sold FOR RESEARCH USE ONLY. Not for use in diagnostic procedures.

This assay was developed by Invivoscribe.

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