How to narrow down chromosomal breakpoints in small and large derivative chromosomes – a new probe set
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Here a new fluorescence in situ hybridization (FISH-) based probe set is presented and its possible applications are highlighted in 34 exemplary clinical cases. The so-called pericentric-ladder-FISH (PCL-FISH) probe set enables a characterization of chromosomal breakpoints especially in small supernumerary marker chromosomes (sSMC), but can also be applied successfully in large inborn or acquired derivative chromosomes. PCL-FISH was established as 24 different chromosome-specific probe sets and can be used in two- up multicolor-FISH approaches. PCL-FISH enables the determination of a chromosomal breakpoint with a resolution between 1 and ∼10 megabasepairs and is based on locus-specific bacterial artificial chromosome (BAC) probes. Results obtained on 29 sSMC cases and five larger derivative chromosomes are presented and discussed. To confirm the reliability of PCL-FISH, eight of the 29 sSMC cases were studied by array-comparative genomic hybridization (aCGH); the used sSMC-specific DNA was obtained by glass-needle based microdissection and DOP-PCR-amplification. Overall, PCL-FISH leads to a better resolution than most FISH-banding approaches and is a good tool to narrow down chromosomal breakpoints.
KeywordsChromosomal breakpoints Fluorescence in situ hybridization (FISH) Pericentric-ladder-FISH (PCL-FISH) Small supernumerary marker chromosomes (sSMC)
The clinical cases were kindly provided by the following colleagues: Australia: J Anderson, Brisbane; Belgium: Dr. J. Vermeesch, Leuven; France: Dr. C. Yardin, Montpellier; Germany: Dr. I. Bartels, Göttingen; Dr. B. Belitz, Berlin; Dr. U. Beudt, Frankfurt; Dr. H.-M. Burow, Oberkirch; Dr. A. Dufke, Tübingen; Dr. G. Gillessen-Kasebach, Lübeck; Dr. D. Huhle, Leipzig; Dr. A. Kuechler, Essen; Dr. T. Martin, Homburg; Dr. A. Meiner, Halle; Dr. D. Mitter, Leipzig; Dr. S. Morlot, Hannover; Dr. A. Ovens-Raeder, München, Dr. G. Schwan, Dortmund; Dr. S. Singer, Tübingen; Dr. S. Spranger, Bremen; Portugal: Dr. J. Melo, Coimbra; Serbia: Dr. G. Josik, Vinca; Turkey: Dr. B. Seher, Ankara; UK: Dr. K. Ren Ong, Birmingham.
Supported in parts by Deutsche Forschungsgemeinschaft (DFG LI 820/22-1), Else Kröner-Fresenius-Stiftung (2011_A42), the Deutscher Akademischer Austauschdienst (DAAD), the Monika-Kutzner-Stiftung and the Stefan-Morsch-Stiftung.
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