Spectral karyotyping SKY is a multi-fluorochrome fluorescence in situ hybridization technique FISH in which all the chromosome pairs are simultaneously visualized in different colors in a single hybridization. SKY determines the unique spectral profile of each chromosome generated by specific combinations of different fluorochromes.
At the present time, SKY can be used to analyze human, mouse, and rat chromosomes. Structural and numerical chromosomal alterations aberrations are the hallmarks of malignant diseases.
Routine cytogenetic analysis based on G-protein couple receptor GPCR techniques provides important information of diagnostic and prognostic relevance both in hematological malignancies and solid tumors. However, detection of chromosomal alterations by this method is complicated by the difficulty in routinely preparing metaphase spreads of sufficient quality and quantity, the clonal heterogeneity of the tumors, and the complexity of the many c Skip to main content Skip to table of contents.
This is often because the complexity of the rearrangement has resulted in the lack of a coherent and recognizable banding pattern. The advent of the various multicolor fluorescence in situ hybridization FISH chromosomal painting techniques 1,1 has greatly improved our ability to identify all marker chromosomes, but these techniques still need some careful planning in rapidly achieving the goal of identifying complex chromosomal rearrangements.
Cancer is a genetic disease. Gene mutations are not only responsible for rare hereditary forms of human cancer, but for the sporadic forms of human malignancies as well.
Many of these specific genetic defects in cancer cells can be visualized as. Many of these specific genetic defects in cancer cells can be visualized as chromosomal aberrations.
Conventional cytogenetic analysis of metaphase chromosomes from human malignancies is a first screening step to identify chromosomal aberrations. Since the introduction of chromosome banding techniques in by Caspersson et al. In these malignancies, specific balanced translocations were identified and have led to the cloning of the genes involved at many breakpoints. These aberrations have proven to be of significant etiologic, diagnostic, prognostic, as well as therapeutic relevance, especially in leukemias.
While cytogenetic analyses have been exceedingly valuable for the description of chromosomal abnormalities in hematologic malignancies and in sarcomas, epithelial cancers were more difficult to study. This is owing, in part, not only to the accessibility of malignant cells and subsequently metaphases for cytogenetic analysis in leukemias, but also to the nature of reciprocal translocations, which provided more immediate entry points for positional cloning efforts.
Multicolor karyotyping of human chromosomes is a valuable tool for defining marker chromosomes as well as small, cryptic or complex chromosome translocations. Although the results obtained are excellent, both procedures require expensive equipment for detection and specialized software for analysis.
In many laboratories, research projects often only occasionally require multicolor karyotyping , and so expensive investment in specialized accessory hardware cannot be justified.
To minimize costs and yet enable color karyotyping using only the most basic equipment, we have developed an alternative technique called color-changing karyotyping CCK , which uses DNA probes labeled with only three fluorescent dyes. Image capture can be accomplished using a conventional digital camera, and image processing can be performed with generic imaging software. An optimized procedure for cytogenetic slide preparation and efficient chromosome hybridization is also detailed.
Coronavirus Resources. Technique: Spectral Karyotyping. Publication Year. Invalid publishing year. Technique Show more Show fewer. Antibody Show more Show fewer. Anti-mouse 9. Organism Show more Show fewer. Homo sapiens Cell Line Show more Show fewer. HEK 1. Nature Research 2. Springer Article Category. Protocol The data presented here and in the literature do not support the use of spectral karyotyping as a primary diagnostic tool in prenatal situations, as the risk of misdiagnosis is still a major concern [ 7 , 8 ].
Yaron et al. In this series, the evaluation of ring chromosomes and non-satellited SMCs benefited greatly from the additional analysis; however, spectral karyotyping did not provide further information for characterization in two cases with bisatellited SMCs [ 7 ]. Therefore, Heng et al. This strategy involves routine G-banding for the initial evaluation, but any marker chromosomes or complex rearrangements thus detected are further characterized by spectral karyotyping and then verified with FISH.
Our findings provide evidence that this strategy is efficient for pre- and postnatal analysis. In this regard, Liehr et al. In the foreseeable future, spectral karyotyping in conjunction with array CGH will continue to be applied not only in constitutional chromosome studies, but also in cancer cytogenetic studies for identification of complex and cryptic rearrangements [ 19 ].
Knowing that chromosomal changes in cancer are often considered a useful reference for therapy design, these chromosomal changes must be elucidated as precisely as possible.
To provide better care for cancer patients, spectral karyotyping in combination with other molecular cytogenetic methodologies such as FISH and array CGH should be useful for supplementing G-banding analysis for the identification of significant prognostic rearrangements. In conclusion, our findings describe the validation of spectral karyotyping in a national-wide commercial cytogenetics laboratory and its successful introduction into the diagnostic arena.
This technology provides a valuable diagnostic tool for establishing the origin of small supernumerary marker chromosomes sSMCs and derivative chromosomal material that cannot be identified with standard techniques. Science , — Hum Genet , — Am J Med Genet , 80 4 — Prenat Diagn , 18 11 — Prenat Diagn , 19 12 — Prenat Diagn , 23 1 — Article PubMed Google Scholar.
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Methods Mol Med , 29— PubMed Google Scholar. Download references. The authors thank Jeff Radcliff Quest Diagnostics for assistance in editorial review of the manuscript. You can also search for this author in PubMed Google Scholar.
Correspondence to Charles M Strom. AA supervised the assay implementation and its performance, supervised molecular cytogenetic studies, drafted and revised the manuscript. BTW reviewed and analyzed the spectral karyotyping data; drafted and finalized the manuscript.
SRW supervised the molecular cytogenetic studies. FZB supervised the molecular cytogenetic studies and made critical comments on the drafted manuscript. LOM collected further clinical information and constructed Table 2. MME reviewed cases referred for spectral karyotyping analysis. PHK reviewed cases referred for spectral karyotyping analysis.
MHH reviewed cases referred for spectral karyotyping analysis. VS carried out the molecular cytogenetic studies. AHS supervised the molecular cytogenetic studies. MSA supervised the molecular cytogenetic studies. BJW reviewed cases referred for spectral karyotyping analysis.
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