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IGH Gene Clonality Assay - ABI Fluorescence Detection
Product Name Cat. No. Size Price Qty.
IGH Gene Clonality Assay - ABI Fluorescence Detection 1-101-0061
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Product Uses
Historically, Immunoglobulin Heavy Chain Gene Rearrangement Assays have been used to:
  1. Identify clonality in atypical lymphoproliferative disorders
  2. Support a differential diagnosis between reactive lesions and hematologic malignancies
  3. Assign presumptive lineage in mature monoclonal lymphoproliferative disorders
  4. Identify tumor-specific markers (IGH gene rearrangements) for post-treatment monitoring
  5. Monitor and evaluate disease recurrence
Overview
InVivoScribe Technologies' assay are standardized PCR-based tests. Each test comes with a Standard Operating Procedure (SOP), an interpretation guide, master mixes, and controls. Master mixes are composed of a buffered magnesium solution, deoxynucleotides, and multiple primers that target the gene segments of interest. Multiple primers are used to ensure a more comprehensive testing approach necessary to reliably identify clonal rearrangements. These tests are complete with the exception of Taq DNA Polymerase, which is not provided. A single thermocycler program and similar detection methods are used within each series of test to improve consistency, reduce human error, and facilitate cross training.

Standard Protocol. Click to read more
Description
The immunoglobulin heavy chain (IGH) gene locus on chromosome 14 (14q32.3) includes 46-52 functional and 30 non-functional variable (VH) gene segments, 27 functional diversity (DH) gene segments, and 6 functional joining (JH) gene segments spread over 1250 kilobases. The most frequently used VH gene segments in normal and malignant B cells belong to the VH3, VH4, and VH1 family, together covering 75-95% of VH usage. The VH gene segments contain three framework regions (FR) and two complementarity determining regions (CDR). The FRs are characterized by their similarity among the various VH segments, whereas the CDRs are highly different even within the same VH family. The CDRs represent the preferred target sequences for somatic hypermutations; however, somatic mutations can also occur in the FRs. Therefore, family-specific primers in the three different 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. In addition to VH-JH rearrangements, incomplete DH-JH rearrangements have been found in mature and immature B cell malignancies. Therefore, DH-JH PCR analysis may be of added value for clonality assessment. The performance characteristics of this assay have been independently determined by a European collaborative study involving 32 diagnostic PCR laboratories (BIOMED-2 Concerted Action) testing hundreds of clinical samples defined according to the WHO classification. Click to read more
Specimen Requirements
This product tests genomic DNA. DNA can be extracted from the following specimens:
  1. 5cc of peripheral blood, bone marrow biopsy, or bone marrow aspirate anti-coagulated with heparin or EDTA; OR,
  2. Minimum 5mm cube of tissue shipped frozen, at room temperature, or on ice in RPMI 1640; OR,
  3. Formalin-fixed paraffin embedded tissue or slides.
Materials Provided
QTY Description
1 Specimen Control Size Ladder - 6FAM
1 IGH Tube A - 6FAM
1 IGH Tube B - 6FAM
1 IGH Tube C - HEX
1 IGH Tube D - HEX
1 IGH Tube E - 6FAM
1 IVS-0000 Polyclonal Control DNA
1 IVS-0008 Clonal Control DNA
1 IVS-0019 Clonal Control DNA
1 IVS-0024 Clonal Control DNA
1 IVS-0030 Clonal Control DNA


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