New concepts to improve resolution and sensitivity of molecular cytogenetic diagnostics by multicolor fluorescence in situ hybridization
✍ Scribed by K. Saracoglu; J. Brown; L. Kearney; S. Uhrig; J. Azofeifa; C. Fauth; M.R. Speicher; R. Eils
- Publisher
- John Wiley and Sons
- Year
- 2001
- Tongue
- English
- Weight
- 507 KB
- Volume
- 44
- Category
- Article
- ISSN
- 0196-4763
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✦ Synopsis
Background: Routine application of multicolor fluorescence in situ hybridization (M-FISH) technology for molecular cytogenetic diagnostics has been hampered by several technical limitations. First, when using chromosome-specific painting probes, there is a limit in cytogenetic resolution of approximately 2-3 Mb, which can mask hidden structural abnormalities that have a significant clinical effect. Second, using whole chromosome painting probes, intrachromosomal rearrangements cannot be detected and the exact localization of breakpoints is often not possible. Methods: We suggest the use of multiplex-labeled region or locus-specific probes in combination with an optimal probe design to improve the sensitivity and resolution of the M-FISH technology. To allow the application of this assay in routine diagnostics, we developed a multipurpose image analysis system.
Results: goldFISH was applied to the study of cryptic translocations in mental retardation patients and to the study of high-resolution breakpoint mapping in non-small cell lung cancer patients. For an individual with mental retardation, who had an apparently normal karyotype by G-banding, we detected an unbalanced translocation involving chromosomes 2 and 7. Conclusions: In combination with optimally designed probe kits, goldFISH overcomes most of the present limitations of the M-FISH technology and results in virtually 100% reliability for detecting interchromosomal and intrachromosomal rearrangements. Cytometry 44:7-15, 2001.