First slide
CytoCell fluorescence in situ hybridisation (FISH) logo.

Product summary

  • Technology FISH
  • Application Haematology
  • Areas of interest MM
  • Region 6p21
    14q32.33
  • Label    
  • Product Code LPH 075 (10 tests)
    LPH 075-S (5 tests)
  • Regulatory Status In vitro diagnostic. This product is intended to be used on Carnoy’s solution (3:1 methanol/acetic acid) fixed haematological samples. Disease information supported by the literature and is not a reflection of the intended purpose of this product.

Chromomaps

Overview

Probe specification

  • IGH, 14q32.33, Green
  • CCND3, 6p21, Red

The IGH/CCND3 Plus product consists of probes labelled in green, proximal to the Constant, and within the Variable segment of the IGH region and CCND3 probes, labelled in red. The CCND3 probe mix contains a 158kb probe telomeric to the CCND3 gene, including the D6S1017 marker and a second probe covering the 244kb region centromeric to the TAF8 gene, including the D6S400 and the D6S1463 markers.

 

Probe information

The CCND3 (cyclin D3) gene is located at 6p21.1 and IGH (immunoglobulin heavy locus) at 14q32.33.

Approximately 50-60% of multiple myeloma (MM) cases are associated with translocations involving IGH and one of several partners including CCND1, NSD2 (WHSC1) and FGFR3, CCND3, MAF or MAFB1.

The t(6;14)(p21;q32) translocation is a recurrent translocation seen in 4% of cases of MM2.

CCND3 has been identified as a putative oncogene that is dysregulated as a consequence of the t(6;14)(p21;q32) translocation2. The translocation appears to be mediated by an error in IgH switch recombination as it has been shown that in KMM-1 cell lines, the translocation disrupts a switch sequence in this region and results in juxtaposition of CCND3 with the IGH promoter, thus elevating the levels of CCND3 expression2. It is thought that this mechanism is similar in all cases of IGH translocation. Most breakpoints appear to be clustered in a region that is fewer than 200kb centromeric to CCND32.

CCND3-IGH translocations are also reported in a variety of other B-cell malignancies, including plasma cell leukaemia, diffuse large B-cell lymphoma (DLBCL) and splenic lymphoma with villous lymphocytes (SLVL)3.

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References

  1. Fonseca et al., Cancer Res 2004;64:1546-58
  2. Shaughnessy et al., Blood 2001;98(1):217-23
  3. Soniki et al., Blood 2001;98(9):2837-44

Recommended protocol for CytoCell haematology FISH

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Sample and slide preparation

Icon representing the sample and slide preparation stage of the fluorescence in situ hybridisation (FISH) protocol.
  • Spot the cell sample onto a glass microscope slide. Allow to dry.
  • Immerse the slide in 2x Saline Sodium Citrate (SSC) for 2 minutes at room temperature (RT) without agitation.
  • Dehydrate in an ethanol series (70%, 85% and 100%), each for 2 minutes at RT.
  • Allow to dry.
Haematology FISH protocol Video Image
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Haematology FISH protocol

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