CAT #: 12070101
T Cell Receptor Gamma Gene Rearrangement Assay 2.0 - ABI Fluorescence Detection
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
- Summary of Explanation of the Test
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 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
- 5 cc of peripheral blood, bone marrow biopsy, or bone marrow aspirate anti-coagulated with heparin or EDTA
- Minimum 5 mm cube of tissue
- 100 ng of genomic DNA
- Formalin-fixed paraffin embedded tissue or slides
This product is sold FOR RESEARCH USE ONLY. Not for use in diagnostic procedures.
This assay was developed by Invivoscribe.
This product is covered by one or more of the following patents and patent applications owned by or exclusively licensed to Invivoscribe, Inc:, United States Patent No. 7,785,783, United States Patent Application Number 10/531,106, European Patent Number EP 1549764B1 and other pending patent applications originating from European Patent Application Numbers 03756746.8 and 047326551.9 (16 countries), Japanese Patent Number JP04708029B2, Japanese Patent Application Number 2006-529437, Brazil Patent Application Number PI0410283.5, Canadian Patent Application Number 2525122, Indian Patent Application Number 5792/DELNP/2005, Mexican Patent Application Number PA/a/2005/012102, Chinese Patent Application Number 200480016603.5, and Korean Patent Application Number 10-2005-7021561.
Use of this product may require nucleic acid amplification methods such as Polymerase Chain Reaction (PCR). Any necessary license to practice amplification methods or to use amplification enzymes or equipment covered by third party patents is the responsibility of the user and no such license is granted by Invivoscribe, Inc, expressly or by implication.
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