Automated fluorescent in situ hybridization (FISH) analysis of t(9;22)(q34;q11) in interphase nuclei

Béla Kajtár, G. Méhes, Thomas Lörch, Linda Deák, Marika Kneifné, Douát Alpár, László Pajor

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Background: For chronic myeloid leukemia, the FISH detection of t(9;22)(q34;q11) in interphase nuclei of peripheral leukocytes is an alternative method to bone marrow karyotyping for monitoring treatment. With automation, several drawbacks of manual analysis may be circumvented. In this article, the capabilities of a commercially available automated image acquisition and analysis system were determined by detecting t(9;22)(q34;q11) in interphase nuclei of peripheral leukocytes. Methods: Three peripheral blood samples of normal adults, 21 samples of CML patients, and one sample of a t(9;22)(q34;q11) positive cell-line were used. Results: Single nuclei with correctly detected signals amounted to 99.6% of nuclei analyzed after exclusion of overlapping nuclei and nuclei with incorrect signal detection. A cut-off value of 0.84 μm was defined to discriminate between translocation positive and negative nuclei based on the shortest distance between signals. Using this value, the false positive rate of the automated analysis for negative samples was 7.0%, whereas that of the manual analysis was 5.8%. Automated and manual results showed strong correlation (R2 = 0.985), the mean difference of results was only 3.7%. Conclusions: A reliable and objective automated analysis of large numbers of cells is possible, avoiding interobserver variability and producing statistically more accurate results than manual evaluation.

Original languageEnglish
Pages (from-to)506-514
Number of pages9
JournalCytometry Part A
Volume69
Issue number6
DOIs
Publication statusPublished - Jun 2006

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Interphase
Fluorescence In Situ Hybridization
Leukocytes
Karyotyping
Observer Variation
Automation
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Cell Count
Bone Marrow
Cell Line
Therapeutics

Keywords

  • Automated analysis
  • BCR-ABL
  • CML
  • FISH

ASJC Scopus subject areas

  • Hematology
  • Cell Biology
  • Pathology and Forensic Medicine
  • Biophysics
  • Endocrinology

Cite this

Automated fluorescent in situ hybridization (FISH) analysis of t(9;22)(q34;q11) in interphase nuclei. / Kajtár, Béla; Méhes, G.; Lörch, Thomas; Deák, Linda; Kneifné, Marika; Alpár, Douát; Pajor, László.

In: Cytometry Part A, Vol. 69, No. 6, 06.2006, p. 506-514.

Research output: Contribution to journalArticle

Kajtár, Béla ; Méhes, G. ; Lörch, Thomas ; Deák, Linda ; Kneifné, Marika ; Alpár, Douát ; Pajor, László. / Automated fluorescent in situ hybridization (FISH) analysis of t(9;22)(q34;q11) in interphase nuclei. In: Cytometry Part A. 2006 ; Vol. 69, No. 6. pp. 506-514.
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