Four Color FISH: A novel four color PDGFRα/KIT/CC4/RET FISH probe set providing coverage of PDGFRα, KIT and RET genes is utilized in this FISH assay to detect PDGFRα/KIT amplification and RET gene rearrangement on the same sample. RET proto-oncogene encodes a receptor tyrosine kinase, which are cell-surface molecules that transduce signals for cell growth and differentiation. PDGFRα or the Platelet- derived growth factor receptor encodes a cell surface tyrosine kinase receptor for members of the platelet derived growth factor family and act as mitogens for cells of mesenchymal origin.
Methodology: A single 4µM FFPE section is hybridized to a novel four color FISH probe set with ORANGE (451Kb) and GREEN (641Kb) fluorophores flanking 5’ and 3’ ends of the RET probe on chromosome 10q11.2; the AQUA labelled PDGFRα/KIT gene on chromosome 4q12 (675Kb) and RED labelled copy control at position 4p12 (501Kb). This assay detects RET gene disruption and PDGFRα/KIT amplification based on evaluation of 50 cells and criteria established in validation studies.