Outside North America 
CAT #: 93090020
IdentiClone® BCL2/JH Translocation Assay - Gel Detection
Intended Use
The IdentiClone® BCL2/JH Translocation Assay is an in vitro diagnostic product intended for PCR-based detection of BCL2/JH t(14;18) gene translocations in patients with suspect lymphoproliferations and can be used to:
- Distinguish lymphoma from benign lymphoid hyperplasia
- Distinguish follicular lymphoma from other B-cell lymphomas that may have a similar appearance
- Monitor and evaluate disease recurrence
Product Details
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Summary and Explanation of the Test
The BCL2 t(14;18) (q32;q21) translocation is found in 80-90% of follicular lymphomas and 30% of diffuse large cell lymphomas; however, this translocation is rarely present in other lymphoproliferative diseases. The t(14;18) brings about a juxtaposition of the BCL2 gene with the immunoglobulin heavy chain (IGH) gene joining segment. This leads to a marked increase in expression of bcl-2 driven by the immunoglobulin heavy chain gene enhancer. The bcl-2 protein inhibits programmed cell death (apoptosis) leading to cell accumulation.
The majority of breakpoints on 18q21-22 occur 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) located 3’ to BCL2 exon 3 (20-25% of the cases). Some breakpoints occur at distant loci and will not be identified by this particular test. Therefore a negative result does not completely exclude the presence of a BCL2/IGH gene rearrangement in the sample.1
Invivoscribe’s IdentiClone assays represent a simple approach to PCR-based clonality testing. These standardized assays were carefully optimized testing positive and negative control samples using multiplex master mixes. Assay development was followed by extensive validation including the testing of more than 400 clinical samples using Revised European/American Lymphoma (REAL) Classification. Testing was performed at more than thirty prominent independent testing centers throughout Europe in a collaborative study known as the BIOMED-2 Concerted Action.1
The gel detection based assays cannot reliably detect clonal populations comprising less than 1% of the total lymphocyte cell population. Always interpret the results of this test in the context of morphologic and other relevant data and do not use alone for a diagnosis of malignancy.
This test kit includes four (4) master mixes. The BCL2/JH translocation master mixes (BCL2/JH Tube A, B, and C) target the joining (J) region of the immunoglobulin heavy (IGH) chain gene and distinct regions of the BCL2 gene. These master mixes are used to detect major breakpoint region (Mbr) and minor cluster region (mcr) of the BCL2 t(14;18) translocations. The fourth master mix, the Specimen Control Size Ladder, targets multiple genes and generates a series of amplicons of approximately 100, 200, 300, 400, and 600 base pairs to ensure that the quality and quantity of input DNA is adequate to yield a valid result. A single thermal cycler program and similar detection methodologies are used with many of our assays which improves consistency and facilitates cross training on a broad range of different assays.
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Principles of the Procedure
Polymerase Chain Reaction (PCR)
PCR assays are routinely used for the identification of chromosome translocations. This test targets the Mbr and mcr regions of the BCL2/JH translocations and amplifies genomic DNA between primers that target the BCL2 gene and the conserved joining (J) regions of the IGH gene (BCL2/JH Tubes A, B, and C). Breakpoints that occur outside the Mbr and mcr regions will not be identified by this particular test. Therefore, a negative result does not completely exclude the presence of a BCL2/JH gene rearrangement in the sample. DNA from a normal lymphocyte population will also generate a negative result.
Gel Detection
Gel electrophoresis, such as agarose gel electrophoresis or non-denaturing polyacrylamide gel electrophoresisis (PAGE), is commonly used to resolve amplicon products based on their size, charge, and conformation. Since DNA is negatively charged, when an electrical potential (voltage) is applied across the gel containing PCR products, the electrical field causes the amplicons to migrate through the gel. Smaller DNA fragments are able to easily migrate through the gel matrix, whereas larger DNA fragments migrate more slowly. This causes a separation of the amplicon products based on size. Ethidium bromide or other DNA intercalating dyes can then be used to stain and detect these products in the gel.
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Specimen Requirements
This assay tests genomic DNA (gDNA) from the following sources:
- 5 cc of peripheral blood, bone marrow biopsy, or bone marrow aspirate anti-coagulated with heparin or EDTA (stored at 2°C to 8°C and shipped at ambient temperature)
- Minimum 5 mm cube of tissue (stored and shipped frozen; or stored and shipped in RPMI 1640 at ambient temperature or on ice)
- 2 µg of gDNA (stored at 2°C to 8°C and shipped at ambient temperature)
- Formalin-fixed paraffin embedded tissue or slides (stored and shipped at ambient temperature)
References
1. Miller, JE et al. (1999) Molecular Diagnostics. 4:101-117.
Disclaimer
This assay is based on the EuroClonality/BIOMED-2 Concerted Action BMH4-CT98-3936.
Legal Notice
Warranty and Liability
Invivoscribe, Inc. (Invivoscribe®) is committed to providing the highest quality products. Invivoscribe® warrants that the products meet or exceed the performance standards described in the Instructions For Use, as to products with such an insert. If a product is covered by product specifications and does not perform as specified, our policy is to replace the product or credit the full purchase price. No other warranties of any kind, expressed or implied, are provided by Invivoscribe®. Invivoscribe® liability shall not exceed the purchase price of the product. Invivoscribe® shall have no liability for direct, indirect, consequential or incidental damages arising from the use, results of use, or inability to use its products; product efficacy under purchaser controlled conditions in purchaser’s laboratory must be established and continually monitored through purchaser defined and controlled processes including but not limited to testing of positive, negative, and blank controls every time a sample is tested. Ordering, acceptance, and use of product constitutes purchaser acceptance of sole responsibility for assuring product efficacy and purchaser agreement to the limitation of liability set forth in this paragraph.
This product is an in vitro diagnostic product is not available for sale or use within North America.
This product is covered by one or more of the following: European Patent Number 1549764, European Patent Number 2418287, European Patent Number 2460889, Japanese Patent Number 4708029, United States Patent 8859748, United States Patent 10280462, and related pending and future applications. All of these patents and applications are licensed exclusively to Invivoscribe®. Additional patents licensed to Invivoscribe covering some of these products apply elsewhere. Many of these products require nucleic acid amplification methods such as Polymerase Chain Reaction (PCR). No license under these patents to use amplification processes or enzymes is conveyed expressly or by implication to the purchaser by the purchase of this product.
IdentiClone® is a registered trademark of Invivoscribe®.
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