Both cytology (brushing and catheter rinse) and histology (biopsies) samples were used for morphological diagnosis. Molecular testing was performed after DNA extraction from sections cut from cell (cytology) or formalin-fixed paraffin-embedded (tissue) blocks, and included a first screening of EGFR (epidermal growth factor receptor) common mutations by the Cobas® technique (Kit ROCHE Cobas DNA Sample Preparation Kit), immunohistochemistry (IHC) for ROS1 (Clone D4D6; Ozyme, Saint-Cyr-l'École, France) and ALK (anaplastic lymphoma kinase, Clone 1A4; Diagomics, Blagnac, France) rearrangements followed by fluorescence in situ hybridisation (FISH) if positive (IQFISH break-apart probe, Dako Omnis, Agilent, Santa Clara, CA, USA)), and in a second time, a hybrid capture NGS, covering a 48-genes panel (Roche Sequencing (Kapa/SeqCap), Roche, Bale, Switzerland; MiSeq DX, Illumina, San Diego, CA, USA). PD-L1 expression was assessed on biopsies and surgically resected specimens using IHC (Clone QR1, Quartett, Diagomics).
Iqfish break apart probe
Manufactured by Agilent Technologies
Sourced in United States
The IQFISH break-apart probe is a molecular biology tool designed to detect specific genetic rearrangements. It is intended for use in fluorescence in situ hybridization (FISH) assays to visualize and analyze chromosomal abnormalities.
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Lab products found in correlation
2 protocols using iqfish break apart probe
1
Comprehensive Molecular Profiling of Lung Cancer
2
Evaluating ALK, ROS1, and MET in NSCLC
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ALK and ROS1 gene rearrangements and MET amplification were evaluated by the standard FISH method. Briefly, unstained sections obtained from FFPE blocks or cytoblocks were incubated with an ALK and ROS1 dual-color probe (IQFISH Break Apart Probe Agilent Technologies, Santa Clara, CA, USA). In each case, at least 100 tumor nuclei were evaluated. Cells were considered positive if a break-apart pattern of orange and green signals, at least one single orange signal, or a combination of both patterns were seen. Tumors with at least 15% of cells with ALK or ROS1 rearrangements were defined as positive. In ambiguous or equivocal cases, ALK or ROS1 immunohistochemical stains (clone D5F3, Ventana, Tucson, AZ, USA and clone D4D6, Cell Signaling, Danvers, MA, USA, respectively) were performed. The presence of MET gene amplification was evaluated using the MET IQFISH Probe with CEP7 (Agilent Technologies, Santa Clara, CA, USA). Amplification was reported in cases with a MET Probe/CEP7 Ratio ≥2 and/or gene copy number ≥5.
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