Highly Multiplexed FISH for in Situ Genomics

Publication date: Available online 9 March 2019Source: The Journal of Molecular DiagnosticsAuthor(s): Maristela L. Onozato, Clarence Yapp, Douglas Richardson, Tilak Sundaresan, Varun Chahal, Jesse Lee, James P. Sullivan, Marisa W. Madden, Hyo Sup Shim, Mathew Liebers, Quan Ho, Shyamala Maheswaran, Daniel A. Haber, Zongli Zheng, Brian Clancy, Hunter L. Elliott, Jochen K. Lennerz, A. John IafrateThe quantification of changes in gene copy number is critical to our understanding of tumor biology and for the clinical management of cancer patients. DNA fluorescence in situ hybridization is the gold standard method to detect copy number alterations, but is limited by the number of genes one can quantify simultaneously. To increase the throughput of this informative technique, a fluorescent “barcode” system for the unique labeling of dozens of genes and an automated image analysis algorithm that enabled their simultaneous hybridization for the quantification of gene copy numbers were devised. We demonstrate the reliability of this multiplex approach on normal human lymphocytes, metaphase spreads of transformed cell lines, and cultured circulating tumor cells. It also opens the door to the development of gene panels for more comprehensive analysis of copy number changes in tissue including the study of heterogeneity and of high throughput clinical assays that could provide rapid quantification of gene copy numbers in samples with limited cellularity such as circulating tumor cells...
Source: The Journal of Molecular Diagnostics - Category: Pathology Source Type: research