Interphase fluorescence in situ hybridization analysis: a study using centromeric probes 7, 8, and 12

Area of Science:

• Cytogenetics
• Molecular Biology
• Oncology

Background:

• Interphase fluorescence in situ hybridization (I-FISH) is increasingly utilized in cytogenetics and clinical diagnostics for detecting chromosomal abnormalities in tumors.
• Standardized performance and quality control data are essential for adopting commercial FISH probes in routine clinical laboratory testing.

Purpose of the Study:

• To evaluate the reliability, reproducibility, and accuracy of commercially available alpha-satellite DNA centromeric probes for I-FISH analysis.
• To assess the feasibility of integrating I-FISH into routine clinical practice.

Main Methods:

• Interphase FISH analysis was conducted using three commercial alpha-satellite chromosome-specific DNA centromeric probes (D7Z1/D7Z2, D8Z2, D12Z3).
• Analysis was performed on bone marrow samples prepared for conventional cytogenetic analysis.
• Signal enumeration in 500 interphase nuclei was conducted by two independent observers for each probe.

Main Results:

• High accuracy rates were observed: 93.92% (+/- 1.3%) for chromosome 7, 93.91% (+/- 1.5%) for chromosome 8, and 92.85% (+/- 1.4%) for chromosome 12.
• The results demonstrated consistent signal enumeration between observers, indicating high reproducibility.
• The study confirmed the reliability and accuracy of the tested I-FISH probes.

Conclusions:

• Interphase FISH utilizing chromosome centromeric probes is a reliable, reproducible, and accurate technique for detecting numerical chromosomal abnormalities.
• With appropriate quality control protocols, I-FISH can be effectively integrated into routine clinical laboratory practice.
• This technique supports advancements in cytogenetic diagnostics and cancer research.