Institute of Human Genetics, Jena University Hospital, Kollegiengasse 10, 07743 Jena, Germany.
J Appl Genet. 2012 Aug;53(3):259-69. doi: 10.1007/s13353-012-0098-9. Epub 2012 Apr 29.
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.
此处提出了一种新的荧光原位杂交（FISH）探针集，并在 34 个示例临床病例中强调了其可能的应用。所谓的着丝粒梯式 FISH（PCL-FISH）探针集特别能够对染色体断裂点进行特征描述，尤其是在小的额外标记染色体（sSMC）中，但也可以成功应用于大的先天或后天衍生染色体。PCL-FISH 建立了 24 种不同的染色体特异性探针集，可用于双色或多色 FISH 方法。PCL-FISH 能够以 1 到 ∼10 兆碱基对的分辨率确定染色体断裂点，其基于基因座特异性细菌人工染色体（BAC）探针。本文呈现并讨论了在 29 个 sSMC 病例和 5 个较大的衍生染色体上获得的结果。为了确认 PCL-FISH 的可靠性，对 29 个 sSMC 病例中的 8 个进行了基于阵列的比较基因组杂交（aCGH）研究；使用的 sSMC 特异性 DNA 通过玻璃针微切割和 DOP-PCR 扩增获得。总体而言，PCL-FISH 比大多数 FISH 带型方法具有更好的分辨率，是缩小染色体断裂点的良好工具。
