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

Product summary

  • Chromosomes
    11
  • Labels    
  • Disease focus CLL
  • Product code CE-LPH 011-S (5 tests) CE-LPH 011 (10 tests)
  • Regulatory status In vitro diagnostic.
Cytocell Catalogue Probe Packaging With Amber Tube
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Probe design

Overview

Probe specification

  • ATM, 11q22.3, Red

  • D11Z1, 11p11.11-q11, 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 protein kinase ATM (ATM serine/threonine kinase) gene at 11q22.3, is frequently deleted in cases of B-cell chronic lymphocytic leukaemia (CLL). ATM is an important checkpoint gene involved in the management of cell damage. 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 pathway1.

B-CLL is the most common leukaemia in adults; its course can vary from very indolent to rapidly progressive. Due to the low mitotic activity of the leukaemic cells in vitro, clonal chromosomal abnormalities are detected in 40-50%2 of cases by conventional cytogenetics using B- cell mitogens, whereas FISH analysis identifies chromosomal aberrations in approximately 80%2 of B-CLLs. Screening for deletions of ATM and/or TP53 is vital to allow informed therapy choices for B-CLL patients, as deletions of TP53 and ATM confer poorer prognosis in this disease4; therefore, the use of FISH has proved to be a powerful tool in both the diagnosis and management of patients with B-CLL2,3,4.

Analysis of the ATM/TP53 interaction in B-CLL has shown that TP53 and ATM play an important role in the proliferation of lymphoid cancer1. It has been shown that ATM enhances the phosphorylation of TP53, should the damage be so great that the cell requires destruction by apoptosis (which is mediated by TP53). Deletion of ATM removes this checkpoint activity and hence activation of TP53. Thus, there is no attempt made to repair, or apoptosis of, damaged cells, despite the presence of TP53. In the absence of ATM, damaged cells are allowed to continue to proliferate5.

 

Intended purpose

The CytoCell® ATM Deletion Probe is a qualitative, non-automated, fluorescence in situ hybridisation (FISH) test used to detect chromosomal deletions in the 11q22.3 region on chromosome 11 in Carnoy’s solution (3:1 methanol/acetic acid) fixed haematologically-derived cell suspensions from patients with confirmed or suspected chronic lymphocytic leukaemia (CLL).

 

Indications for use

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

 

Limitations

This device is designed to detect genomic losses larger than the region covered by the red clone in this probe set, which includes the ATM region. Genomic losses outside this region or partial losses of this region may not be detected with this device.

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 use.

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. Stankovic, et al. Blood. 2004;103(1):291-300.
  2. Dohner, et al. N Eng J Med. 2000;343:1910-1916.
  3. Zent, et al. Blood. 2010;115(21):4154-4155.
  4. Rossi, et al. Blood. 2013;121(8):1403-1412.
  5. Khanna, et al. Nature Genetics. 1998;20(4):398-400.
  6. Arsham, MS., Barch, MJ. and Lawce HJ. (eds.) (2017) The AGT Cytogenetics Laboratory Manual. New Jersey: John Wiley & Sons Inc.
  7. 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.
  8. 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.