b'Gel & Capillary | RUO AssaysGel and CapillaryTCRG Gene Clonality AssaysResearch Use Only (RUO) AssayTCRG Gene Clonality AssaysAssay Use smaller part of B NHLs. In addition, the TRG gene contains a limited number of Vand Jsegments such that the amplification of all major TRG Gene Clonality Assays are useful for studies involving: V-Jcombinations is possible with four Vand two Jprimers. dentification of clonal T-cell populations highly suggestive ofI This standardized multiplex PCR assay detects the vast majority T-cell malignancies of clonal TRG gene rearrangements using only two multiplex Lineage determination of leukemias and lymphomas master mixes. 1The potential risk of false-positive results, due to Monitoring and evaluation of disease recurrence overinterpretation of minor clonal peaks, can be minimized by the combined use of heteroduplex analysis and differential fluorescence Detection and assessment of residual disease detection, and by interpreting results within their clinical context. 1,2Evaluation of new research and methods in malignancy studies The detection rate of clonal TRG gene rearrangements using this assay is exceptionally high. 1 Summary and Explanation of the TestThis assay tests all variable (V ) regions 1-11 of the TRG (formerlySpecimen Requirementsknown as TCRG) gene Master mix tubes A and B target conserved regions within the variable (V ) and joining (J ) regions that flank5 mL of peripheral blood, bone marrow biopsy, or bone marrow the unique, hypervariable, antigen-binding, complementarity determining region 3 (CDR3). Tube A contains two Vprimers and twoaspirate anti-coagulated with heparin or EDTA; or,J primers. Tube B contains two Vprimers and two J primers. PositiveMinimum 5 mm cube of tissue; or,and negative controls, as well as a Specimen Control Size Ladder2 g of genomic DNA; or,Master Mix are included. PCR products can be analyzed by capillary electrophoresis or heteroduplex analysis. Clonality is indicated if anyFormalin-fixed, paraffin-embedded (FFPE) tissue or slides.one of the master mixes generates clonal products.ReferenceBackground1.K Beldjord et al., Leukemia 17: 2289-2292 (2003).The T-cell receptor gamma (TRG, formerly known as TCRG) chain2. JJM van Dongen et al., Leukemia 17: 2257-2317 (2003).locus spans 128 kb on chromosome 7 (7p14). Rearrangement of the variable (V ) and joining (J ) genetic segments of the TRG locus result in V-J products of unique length and sequence. The TRG locus does not contain diversity (D) segments. 2TRG is a preferential target for clonality analyses since it is rearranged in greater than 90% of T-ALL, T-large granular lymphocyte (LGL), and T-PLL, in 50-75% of peripheral T-NHL and mycosis fungoides, but not NK cell proliferations. It is also rearranged in a major part (60%) of B-lineage ALLs and in a much This assay is based on the EuroClonality/BIOMED-2 Concerted Action BMH4-CT98-3936.Figure Legend: Depicted is a simple representation of the organization of the T-cell receptor gamma chain gene on chromosome 7. Black arrows represent the relative positions of primers that target the variable (V) regions, and the downstream joining (J) gene segments. The amplicon products generated from each of these regions can be differentially detected when fluorescent primer sets are used with capillary electrophoresis instruments that employ differential fluorescence detection.114'