Abstract
Spectral FISH is a powerful multiplex fluorescence in situ hybridization (FISH) technique requiring only a single hybridization to target disease-specific multiple chromosomal aberrations in interphase nuclei, using combinatorial fluorescence and digital imaging microscopy developed for the ASIā¢ Spectral Karyotyping System. In oncology, it becomes a particularly useful tool to diagnosis and monitor clonal aberrations not readily amenable to PCR technology, including aneuploidy or recurrent deletions of variable chromosomal regions, such as del(5q), del(6q), or del(7q). Multiple aneuploidies are common in both hematologic malignancies and solid tumors, with particularly frequent involvement of chromosomes 4, 5, 6, 7, 8, 9, 10, 11, 13, 14, 18, 21, X, and Y. To assess ploidy status in low or non-mitotic cells from various neoplastic disorders, including breast cancer, lymphoma. acute rymphoblastic leukemia (ALL), acute myeloid leukemia, and cells recovered from bladder washings suspicious for recurrence of bladder cancer, we developed an initial Spectral FISH oncology panel of six oentromeric probes for chromosomes 7, 8, 9, 10, X, and Y, using unique two-dye combinations of four fluorophores: Cy5.5, Spectrum Green, Spectrum Orange, and Spectrum Red. Spectral FISH readily identified clonal aberrations in all recurrent bladder cancer cases tested and identified MRD in hyperdiploid ALL, providing proof of concept. Validations of the Spectral FISH assay were performed by classic cylogenetics, when mitotic cells were available, or by standard FISH analyses. Advantages of Spectral FISH include increased sensitivity by surveying multiple genetic targets in specimens with limited cell numbers for analysis, identifying aberrations in poor growth/no growth specimens, and Die ability to detect specific genetic aberrations at the single cell level to assess MRD or early stages of recurrent disease. To obviate the limitations of low cell numbers, spectral FISH requires only a single hybridization, while allowing simultaneous detection of multiple numerical changes, with the ability to select combinations of informative probes highly associated with, or characteristic for a specific malignancy. Disadvantages include labor intensive screening, and interpretative challenges associated wilh signal overlap in highly polyploid samples, and focal plane distortions resulting in false positives or negatives. Spectral FISH, as a sensitive MRD assay, has significant potential clinical application, allowing early detection of new or re-emerging clones, and earlier therapeutic intervention.
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Murata-Collins, J., Zhang, F., Tcheurekdiian, L. et al. Interphase Spectral FISH: Tailoring a diagnostic and minimal residual disease (MRD) assay for oncology. Genet Med 2, 94 (2000). https://doi.org/10.1097/00125817-200001000-00155
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DOI: https://doi.org/10.1097/00125817-200001000-00155