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How to narrow down chromosomal breakpoints in small and large derivative chromosomes – a new probe set

  • Human Genetics • Original Paper
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Abstract

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.

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Acknowledgments

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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Authors and Affiliations

  1. Institute of Human Genetics, Jena University Hospital, Kollegiengasse 10, 07743, Jena, Germany

    Ahmed B. Hamid, Katharina Kreskowski, Anja Weise, Nadezda Kosayakova, Kristin Mrasek, Martin Voigt, Roberta Santos Guilherme, Rebecca Wagner, Thomas Liehr & Elisabeth Klein

  2. Chambon Laboratory for Molecular diagnostics (member of the synlab Czech laboratory group), Prague, Czech Republic

    David Hardekopf & Sona Pekova

  3. Laboratory of Morphology and Function of Cell structure, Institute of Cytology and Genetics, Russian Academy of Sciences, Siberian Branch, Lavrentiev Ave. 10, 630090, Novosibirsk, Russian Federation

    Tatyana Karamysheva

  4. Institut für Humangenetik, Postfach, 07740, Jena, Germany

    Thomas Liehr

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  1. Ahmed B. Hamid
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  2. Katharina Kreskowski
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  3. Anja Weise
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  4. Nadezda Kosayakova
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  7. Roberta Santos Guilherme
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  8. Rebecca Wagner
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  9. David Hardekopf
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  11. Tatyana Karamysheva
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  12. Thomas Liehr
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  13. Elisabeth Klein
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Correspondence to Thomas Liehr.

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Hamid, A.B., Kreskowski, K., Weise, A. et al. How to narrow down chromosomal breakpoints in small and large derivative chromosomes – a new probe set. J Appl Genetics 53, 259–269 (2012). https://doi.org/10.1007/s13353-012-0098-9

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  • DOI: https://doi.org/10.1007/s13353-012-0098-9

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