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

Product summary

  • Technology FISH
  • Application Haematology
  • Areas of interest CLL
  • Region 6p11.1-q11.1 / 6q23.3
    11p11.1-q11.1 / 11q22.3
    12p11.1-q11.1
    13q14.2-q14.3 / 13q34
    17p13.1 / 17p11.1-q11.1
  • Label    
  • Product Code LPH 087 (5x10 tests)
    LPH 087-S (5x5 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.

Overview

A selection of haematology probes and an alpha-satellite probe for Chronic Lymphocytic Leukaemia (CLL).

Probes

Alpha Satellite 12 Plus for CLL

Probe specification

  • D12Z3, 12p11.1-q11.1, Red

The Alpha Satellite 12 Plus Probe is a repeat sequence probe, labelled in red, which recognises the centromeric repeat sequence D12Z3.

Probe information

Trisomy 12 is a recurrent abnormality in CLL, seen in 20% of cases1 and often appears as the unique cytogenetic aberration (40-60% of cases with trisomy 12)2. Patients with trisomy 12 are classified as low-risk in the absence of any other genetic lesions3. This product is also available in 5 (LPH 069-S) and 10 (LPH 069) test kit sizes and has been optimised for overnight hybridisation.

13q14.3

Probe specification

  • 13q14.2-q14.3, Red
  • 13qter, 13q34, Green

The 13q14.2-q14.3 probe, labelled in red, covers the D13S319 and D13S25 markers. The 13qter subtelomere specific probe (clone 163C9), labelled in green, allows identification of chromosome 13 and acts as a control probe.

Probe information

Deletions affecting 13q14 are the most frequent structural genetic aberrations in CLL3,4,5. This region is found to be heterozygously deleted in 30-60% and homozygously deleted in 10-20% of CLL patients6. Patients with 13q14 deletions are classified as very low risk, in the absence of any other genetic lesions3.

 

P53 (TP53) (17p13)

Probe specification

  • P53, 17p13.1, Red
  • D17Z1, 17p11.1-q11.1, Green

The P53 probe mix consists of a 161kb probe, labelled in red that covers the whole P53 (TP53) gene and the flanking regions. The probe mix also contains a control probe for the 17 centromere (D17Z1) that is labelled in green.

Probe information

The TP53 (tumor protein p53) gene at 17p13 is one of most important tumour suppressor genes; it acts as a potent transcription factor with fundamental role in the maintenance of genetic stability. Loss of TP53 is reported in 10% of patients with CLL, and is considered to be the poorest prognostic marker3,7.

ATM (11q22.3)

Probe specification

  • ATM, 11q22.3, Red
  • D11Z1, 11p11.1-q11.1, Green

The ATM probe is 182kb, labelled in red, and covers the telomeric end of the NPAT gene and the centromeric end of the ATM gene to just beyond the D11S3347 marker. The probe mix also contains a control probe for the 11 centromere (D11Z1) labelled in green.

Probe information

The ATM (ATM serine/threonine kinase) gene at 11q22.3 is an important checkpoint gene involved in the management of cell damage and its function is to assess the level of DNA damage in the cell and attempt repair by phosphorylating key substrates involved in the DNA damage response pathway8. Loss of ATM is reported in 18% of patients with CLL, and is considered a poor prognostic marker in CLL9.

MYB (6q23.3)

Probe specification

  • MYB, 6q23.3, Red
  • D6Z1, 6p11.1-q11.1, Green

The MYB probe mix consists of a 183kb probe, labelled in red that covers the entire MYB gene and a region telomeric to the gene that includes a centromeric part of the AHI1 gene. This probe mix also contains a control probe for the 6 centromere (D6Z1) labelled in green.

Probe information

Deletions of chromosome 6q are recurrent in CLL. The MYB (MYB protooncogene, transcription factor) gene is essential in haematopoietic cell proliferation and differentiation10,11. It is located in band 6q23.3 and is provided as a marker for 6q deletion.

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References

  1. Swerdlow et al., (eds,) WHO Classification of Tumours of Haematopoietic and Lymphoid Tissue, Lyon, France, 4th edition, IARC,2017
  2. Puiggros et al., Biomed Res Int 2014;1-13
  3. Rossi et al., Blood 2013;121(8):1403-1412
  4. Juliusson G et al., N Eng J Med 1990;323:720-4
  5. Kasar et al., Nature Communications 2015;6:1-12
  6. Hammarsund M et al., FEBS Letters 2004;556:75-80
  7. Baliakas P, et al., Leukemia. 2014;(April):1-8
  8. Stankovic et al., Blood 2004;103(1):291-300
  9. Dohner et al., N Eng J Med 2000;343:1910-1916
  10. Clappier et al., Blood 2007;110(4):1251-1261
  11. Stilgenbauer et al., Leukemia,1999;13:1331-1334

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.
Haematology FISH protocol Video Image
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Haematology FISH protocol

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