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Related Concept Videos

FISH - Fluorescent In-situ Hybridization02:07

FISH - Fluorescent In-situ Hybridization

Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...

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Associated Chromosome Trap for Identifying Long-range DNA Interactions
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A multicenter investigation with D-FISH BCR/ABL1 probes.

G Dewald1, R Stallard, A Alsaadi

  • 1Cytogenetics Laboratory, Mayo Clinic, Rochester, MN 55905, USA.

Cancer Genetics and Cytogenetics
|January 20, 2000
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Summary

This study evaluated a new fluorescence in situ hybridization (FISH) method for chronic myeloid leukemia detection. The D-FISH strategy showed promise, with no false negatives but some false positives, highlighting areas for improved quality assurance.

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Area of Science:

  • Hematology
  • Molecular Diagnostics
  • Cytogenetics

Background:

  • Chronic myeloid leukemia (CML) diagnosis relies on detecting the Philadelphia chromosome.
  • Fluorescence in situ hybridization (FISH) is a key molecular diagnostic tool for CML.
  • Standardization and validation of FISH methods are crucial for reliable clinical practice.

Purpose of the Study:

  • To evaluate a novel dual-color fluorescence in situ hybridization (D-FISH) strategy for CML detection.
  • To assess the clinical sensitivity, precision, and workload of the D-FISH method.
  • To provide recommendations for training and quality assurance in D-FISH implementation.

Main Methods:

  • Twenty-eight laboratories participated in a three-part study using bcr/abl1 D-FISH probes on bone marrow specimens.
  • Laboratories trained on known samples, then blindly analyzed normal and abnormal specimens.
  • Serial dilutions with varying percentages of abnormal nuclei were tested by laboratories and experts.

Main Results:

  • The D-FISH strategy identified no false-negative cases.
  • Fifteen false-positive cases were reported, with abnormal nuclei ranging from 1-6.6%.
  • Participant results for serial dilutions approximated expected values; expert results showed higher precision.

Conclusions:

  • The evaluated D-FISH strategy demonstrates potential for CML diagnosis with high sensitivity.
  • False-positive results warrant attention for optimizing the normal cutoff and quality control.
  • Further training and quality assurance are recommended for robust clinical application of D-FISH.