b'Gel & Capillary | RUO AssaysGel and CapillaryBCL2/JH t(14;18) Translocation AssayResearch Use Only (RUO) AssayBCL2/JH t(14;18) Translocation AssayAssay Use Background The BCL2/J Ht(14;18) Translocation Assay is useful for studies involving: The IGH-BCL2 t(14;18)(q32;q21) translocation is found in 80-90% of follicular lymphomas and in 30% of diffuse large cell lymphomas. 1,2 Monitoring and evaluation of follicular lymphomas and otherThe translocation is rarely present in other lymphoproliferativeB-cell lymphomas diseases. 2The t(14;18) brings about juxtaposition of BCL2 with the Distinguishing lymphoma from benign lymphoid hyperplasia Ig heavy chain joining segment. This leads to a marked increase in expression of BCL2 driven by the Ig heavy chain gene enhancer. 1 Distinguishing follicular lymphoma from other B-cell lymphomas The BCL2 protein inhibits programmed cell death (apoptosis) leading that may have a similar appearance to cell accumulation. 2The majority of breakpoints on 18q21-22 occur Monitoring and evaluation of disease recurrence within the major breakpoint region (MBR) in the 3 untranslated region of exon 3 (60-70% of the cases), and the minor cluster region (mcr) Detection and assessment of residual disease located 3 to BCL2 exon 3 (20-25% of the cases). 1Some breakpoints Evaluation of new research and methods in malignancy studies occur at distant loci and will not be identified by this particular test. 2Therefore, a negative result does not completely exclude the presence of a IGH-BCL2 gene rearrangement in the sample. In comparison, Summary and Explanation of the TestSouthern blot analysis requires 1-2 weeks, is significantly less sensitive, Five master mixes are included in this assay kit. Two master mixesand has more restrictive Specimen requirements.target BCL2 major break point (MBR) translocations and two target BCL2 minor cluster region (mcr) translocations. An Amplification Control Master Mix is also included to ensure the quality and quantitySpecimen Requirementsof sample DNA. Positive and negative controls are also included. This5 mL of peripheral blood, bone marrow biopsy, or bone marrow assay can be run either in a standard or nested assay format.aspirate anti-coagulated with heparin or EDTA; or,Using the standard method, the limit of detection is one cell in oneMinimum 5 mm cube of tissue; or,hundred normal cells. The nested method has a limit of detection of3 g of genomic DNA; or,one t(14;18) positive cell in a background of ten thousand normal cells. PCR products can be analyzed by standard gel electrophoresis withFormalin-fixed, paraffin-embedded (FFPE) tissue or slides.ethidium bromide staining. A BCL2 translocation is indicated if just one of the 2 ndround master mixes (mixes ending in b) generates product(s)Referencewithin the valid size range.1.MS Lee et al., Science 237: 175-178 (1987).2. M Crescenzi et al., Proc. Natl. Acad. Sci. USA 85: 4869-4873 (1988).Figure Legend: Simplified view of the genomic organization of theBCL2 GeneBCL2 and IGH genes on chromosomes 18 and 14, respectively. YellowChromosome 18q21boxes represent the exon regions of the BCL2 gene. Exons of the immunoglobulin heavy chain gene are represented in other colors. The solid black lines represents intron regions, which have been leftHeavy Chain Geneincompletely spliced to assist in demarcation of the exon segments.Chromosome 14q32MBR and mcr type t(14;18) translocations are shown in the lower portions of the figure with the relative positions of primers and the sizeBCL2/IgH Fusion Geneof the amplicons generated from the positive control DNAs indicated. MBR J HBreakpointsBCL2/IgH Fusion Genemcr J HBreakpoints120'