Hamid Ahmed B, Fan Xiaobo, Kosyakova Nadezda, Radhakrishnan Gopakumar, Liehr Thomas, Karamysheva Tatyana
Jena University Hospital, Institute of Human Genetics, Friedrich Schiller University, Kollegiengasse 10, Jena, D-07743, Germany.
Methods Mol Biol. 2015;1227:279-87. doi: 10.1007/978-1-4939-1652-8_14.
Fluorescence in situ hybridization (FISH) and/or array-comparative genomic hybridization (aCGH) performed after initial banding cytogenetics is still the gold standard for detection of chromosomal rearrangements. Although aCGH provides a higher resolution, FISH has two main advantages over the array-based approaches: (1) it can be applied to characterize balanced as well as unbalanced rearrangements, whereas aCGH is restricted to unbalanced ones, and (2) chromosomal aberrations present in low level or complex mosaics can be characterized by FISH without any problems, while aCGH requires presence of over 50 % of aberrant cells in the sample for detection. Recently, a new FISH-based probe set was presented: the so-called pericentric-ladder-FISH (PCL-FISH) that enables characterization of chromosomal breakpoints especially in mosaic small supernumerary marker chromosomes (sSMC). It can also be applied on large inborn or acquired derivative chromosomes. The main feature of this set is that the probes are applied in a chromosome-specific manner and they align along the chromosome in average intervals of ten megabasepairs. Hence PCL-FISH provides denser coverage and a more precise anchorage on the human DNA-sequence than most other FISH-banding approaches.
在最初的显带细胞遗传学之后进行的荧光原位杂交（FISH）和/或阵列比较基因组杂交（aCGH）仍然是检测染色体重排的金标准。尽管aCGH提供了更高的分辨率，但与基于阵列的方法相比，FISH有两个主要优点：（1）它可用于表征平衡和不平衡重排，而aCGH仅限于不平衡重排；（2）低水平或复杂嵌合体中存在的染色体畸变可用FISH毫无问题地表征，而aCGH检测样本中需要存在超过50%的异常细胞。最近，提出了一种基于FISH的新探针组：所谓的着丝粒阶梯FISH（PCL-FISH），它能够表征染色体断点，特别是在嵌合小额外标记染色体（sSMC）中。它也可应用于大的先天性或获得性衍生染色体。该探针组的主要特点是探针以染色体特异性方式应用，并且它们沿着染色体以平均每十个兆碱基对的间隔排列。因此，与大多数其他FISH显带方法相比，PCL-FISH在人类DNA序列上提供了更密集的覆盖和更精确的定位。
