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

Product summary

  • Chromosomes
    14
  • Labels    
  • Disease focus ALL, CLL, Lymphoma, MM
  • Product code CE-LPH 070-S (5 tests) CE-LPH 070 (10 tests)
  • Regulatory status In vitro diagnostic.
Cytocell Catalogue Probe Packaging With Amber Tube
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Probe design

Overview

Probe specification

  • IGHC, 14q32.33, Red

  • IGHV, 14q32.33, Green

The IGH probe mix consists of a 393kb probe, labelled in red, centromeric to the Constant region of the gene and two green probes (324kb and 289kb), within the Variable segment of the gene.

Probe information

Recurrent rearrangements involving the IGH (immunoglobulin heavy locus) at 14q32.33 with a wide range of partner genes are observed in lymphomas and other haematological malignancies.1,2 The IGH locus is a frequent partner in chromosomal translocations involving oncogenes as a consequence of normal immunoglobulin gene recombination processes.1,2

A t(8;14)(q24;q32) translocation involving IGH and MYC is characteristic of Burkitt lymphoma and is also observed in a subset of diffuse large B-cell lymphoma (DLBCL) cases.3,4 Other recurrent IGH rearrangements in B-cell lymphomas include t(14;18)(q32;q21) involving BCL2, commonly identified in follicular lymphoma and DLBCL,5 and t(11;14)(q13;q32) involving CCND1, which is characteristic of mantle cell lymphoma.2,6

In multiple myeloma, recurrent IGH translocations include t(4;14)(p16;q32) involving FGFR3 and NSD2, t(6;14)(p21;q32) involving CCND3, t(11;14)(q13;q32) involving CCND1, t(14;16)(q32;q23) involving MAF, and t(14;20)(q32;q12) involving MAFB.7,8

IGH rearrangements have also been reported in chronic lymphocytic leukaemia (CLL), acute lymphoblastic leukaemia (ALL) and other B-cell lymphoproliferative disorders.2

The breakapart design of this probe set enables detection of rearrangements involving the IGH Locus irrespective of the partner gene or chromosome involved.

 

Intended purpose

The CytoCell® IGH Plus Breakapart Probe is a qualitative, non-automated, fluorescence in situ hybridisation (FISH) test used to detect chromosomal rearrangements in the 14q32.33 region on chromosome 14 in Carnoy’s solution (3:1 methanol/acetic acid) fixed haematologically-derived cell suspensions from patients with confirmed or suspected acute lymphoblastic leukaemia (ALL), chronic lymphocytic leukaemia (CLL), multiple myeloma (MM) or non-Hodgkin lymphoma (NHL).

 

Indications for use

This product is designed as an adjunct to other clinical and histopathological tests in recognised diagnostic and clinical care pathways, where knowledge of IGH rearrangement status would be important for clinical management.

 

Limitations

This device is designed to detect rearrangements with breakpoints in the region bounded by the red and green clones in this probe set, which includes the IGH region. Breakpoints outside this region or variant rearrangements wholly contained within this region may not be detected with this product.

This device is not intended for: use as a stand-alone diagnostic, use as a companion diagnostic, prenatal testing, population-based screening, near-patient testing, or self-testing.

This device has not been validated for sample types, disease types, or purposes outside of those stated in the intended purpose.

It is intended as an adjunct to other diagnostic laboratory tests and therapeutic action should not be initiated on the basis of the FISH result alone.

Reporting and interpretation of FISH results should be performed by suitably qualified staff, consistent with professional standards of practice, and should take into consideration other relevant test results, clinical and diagnostic information.

This device is intended for laboratory professional use only.

Failure to adhere to the protocol may affect the performance and lead to false positive/negative results.

Product documentation

Recommended protocol for CytoCell haematology FISH

Select a protocol step to view:

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.

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References

  1. Gozzetti A, et al. Identification of Novel Cryptic Translocations Involving IGH in B-Cell Non-Hodgkin's Lymphomas. Cancer Res. 2002;62(19):5523-5527.
  2. Alaggio R, Amador C, Anagnostopoulos I, et al. The 5th edition of the World Health Organization Classification of Haematolymphoid Tumours: Lymphoid Neoplasms. Leukemia. 2022;36:1720-1748.
  3. Ferry JA. Burkitt lymphoma: clinicopathologic features and differential diagnosis. Oncologist. 2006;11(4):375-383.
  4. Li S, et al. MYC/BCL2 double-hit high-grade B-cell lymphoma. Mod Pathol. 2012;25(1):145-156.
  5. Snuderl M, et al. BCL2 rearrangements in follicular lymphoma and diffuse large B-cell lymphoma. Am J Surg Pathol. 2010;34(3):327-340.
  6. Campo E, Rule S. Mantle cell lymphoma: evolving management strategies. Blood 2014,48-5.
  7. Bergsagel PL, et al. Promiscuous translocations into immunoglobulin heavy chain switch regions in multiple myeloma. Proc Natl Acad Sci USA. 1996;93(24):13931-13936.
  8. Sawyer JR. The prognostic significance of cytogenetics and molecular profiling in multiple myeloma. Cancer Genet. 2011;204(1):3-12.
  9. Arsham, MS., Barch, MJ. and Lawce HJ. (eds.) (2017) The AGT Cytogenetics Laboratory Manual. New Jersey: John Wiley & Sons Inc.
  10. Mascarello JT, Hirsch B, Kearney HM, et al. Section E9 of the American College of Medical Genetics technical standards and guidelines: fluorescence in situ hybridization. Genet Med. 2011;13(7):667-675.
  11. Wiktor AE, Dyke DLV, Stupca PJ, Ketterling RP, Thorland EC, Shearer BM, Fink SR, Stockero KJ, Majorowicz JR, Dewald GW. Preclinical validation of fluorescence in situ hybridization assays for clinical practice. Genetics in Medicine. 2006;8(1):16–23.
  12. Wolff DJ, Bagg A, Cooley LD et al. Guidance for Fluorescence in Situ Hybridisation Testing in Hematologic Disorders. J Med Diagnost. 2007;9(2):134-143