b'Gel & Capillary | RUO AssaysGel and CapillaryIGH Gene Clonality AssaysResearch Use Only (RUO) AssayIGH Gene Clonality AssaysAssay UseIGH Gene Clonality Assays are useful for studies involving: 7595% of V Husage. The V Hgene segments contain three framework regions (FR) and two complementarity determining regions (CDR).Identification of clonal B-cell populations highly suggestive of B-cell malignancies The FRs are characterized by their similarity among the various V HLineage determination of leukemias and lymphomas segments, whereas the CDRs are highly different even within the same V Hfamily. The CDRs represent the preferred target sequences for Monitoring and evaluation of disease recurrence somatic hypermutations; however, somatic mutations can also occur Detection and assessment of residual disease in the FRs. Therefore, family-specific primers in the three different Evaluation of new research and methods in malignancy studies FRs were designed to increase the detection rate of clonal IGH B-cell populations and decrease the occurrence of false-negative results due to somatic hypermutation in primer binding sites. 1In addition to Summary and Explanation of the TestV H -J Hrearrangements, incomplete D H -J Hrearrangements have been found in mature and immature B-cell malignancies. Therefore, D H -J HFive master mixes target conserved regions within the variable (V H ),PCR analysis may be of added value for clonality assessment. 2diversity (D H ), and the joining (J H ) regions that flank the unique hypervariable, antigen-binding, complementarity determining region 3 (CDR3). Tube A contains six framework region 1 (FR1) primers and a consensus J Hregion primer. Tube B contains seven framework region 2 (FR2) primers and a consensus J Hprimer. TubeSpecimen RequirementsC contains seven framework region 3 (FR1) primers and a consensus5 mL of peripheral blood, bone marrow biopsy, or bone marrow J Hprimer. Tube D contains six D Hregion primers and a consensus J Hregion primer. Tube E contains a D H 7 region primer and a consensusaspirate anti-coagulated with heparin or EDTA; or,J Hprimer. Positive and negative controls, as well as the SpecimenMinimum 5 mm cube of tissue; or,Control Size Ladder Master Mix are included. PCR products can3 g of genomic DNA; or,be analyzed by capillary electrophoresis or heteroduplex analysis. Clonality is indicated if any one of the master mixes generates aFormalin-fixed, paraffin-embedded (FFPE) tissue or slides.clonal product.ReferenceBackground1.M Hummel et al., Leukemia 17:2266-2272 (2003).The immunoglobulin heavy chain (IGH) gene locus on chromosome2. AW Langerak et al., Leukemia 17:2272-2275 (2003).14 (14q32.33, formerly 14q32.3) includes 46-52 functional and 303. JJM van Dongen et al., Leukemia 17:2257-2317 (2003).nonfunctional variable (V H ), 27 functional diversity (D H ), and 6 functional joining (J H ) gene segments spread over 1250 kilobases. 1,2The most frequently used V Hgene segments in normal and malignant B cells belong to the V H 3, V H 4, and V H 1 family, together covering This assay is based on the EuroClonality/BIOMED-2 Concerted Action BMH4-CT98-3936.Figure Legend: Simple representation of the organization of a rearranged immunoglobulin heavy chain gene on chromosome 14. Black arrows represent the relative positions of primers that target the conserved framework (FR1-3) and diversity (D H 1-7) regions, and the downstream consensus J Hgene 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.100'