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Updated: Jul 3, 2026
Ultra-Fast Amplicon-Based Next-Generation Sequencing in Non-Squamous Non-Small Cell Lung Cancer
Published on: September 8, 2023
Gene fusions in non-small cell lung carcinoma (NSCLC): Expert perspectives and practical considerations for routine
Beatriz Bellosillo1, Javier Freire2, Ihab Abdulkader Nallib3
1Department of Pathology, Hospital del Mar, Hospital del Mar Research Institute, Barcelona, Spain; Centro de Investigación Biomédica en Red de Oncología (CIBERONC-ISCIII), Madrid, Spain; Department of Medicine and Life Sicences, Universitat Pompeu Fabra, Spain.
Abstract:
Non-small cell lung carcinoma (NSCLC) encompasses a diverse range of molecular subtypes that require precise identification to guide therapy selection. Among these molecular alterations, gene fusions involving ALK, ROS1, RET, and NTRK present significant challenges for reliable detection in routine clinical practice, particularly due to formalin fixation and the limited availability of nucleic acids, which often hinder molecular analyses. This article highlights key considerations spanning the pre-analytical, analytical, and post-analytical stages, emphasizing the critical need to standardize the entire workflow to ensure high-quality results in gene fusion detection through molecular biology techniques. Particular attention is given to next-generation sequencing (NGS) approaches, with RNA-based NGS strategies being highlighted for their superior sensitivity and accuracy in identifying gene fusions. In conclusion, this work consolidates practical recommendations for integrating optimized gene fusion detection into routine clinical workflows. These include guidance on assay design, quality control measures, and method validation, to ensure the delivery of reliable results to support personalized treatment strategies in NSCLC.
Insights
Accurate detection of gene fusions in non-small cell lung carcinoma (NSCLC) is crucial for personalized therapy. Standardizing workflows and utilizing RNA-based next-generation sequencing (NGS) improves the reliability of identifying these critical molecular alterations.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Non-small cell lung carcinoma (NSCLC) has diverse molecular subtypes requiring precise identification for targeted therapy.
- Gene fusions (e.g., ALK, ROS1, RET, NTRK) are key therapeutic targets but challenging to detect reliably.
- Formalin fixation and limited nucleic acid availability hinder routine molecular analysis of gene fusions.
Purpose of the Study:
- To highlight critical considerations for high-quality gene fusion detection in NSCLC.
- To emphasize the need for standardized pre-analytical, analytical, and post-analytical workflows.
- To provide practical recommendations for integrating optimized gene fusion detection into clinical practice.
Main Methods:
- Review of pre-analytical, analytical, and post-analytical stages in molecular diagnostics.
- Focus on next-generation sequencing (NGS) techniques, particularly RNA-based NGS.
- Consolidation of practical recommendations for assay design, quality control, and validation.
Main Results:
- Standardization across all workflow stages is essential for reliable gene fusion detection.
- RNA-based NGS demonstrates superior sensitivity and accuracy for identifying gene fusions compared to other methods.
- Optimized workflows enhance the delivery of dependable results for personalized NSCLC treatment.
Conclusions:
- Implementing standardized, high-quality gene fusion detection is vital for effective NSCLC therapy selection.
- RNA-based NGS is a recommended approach for sensitive and accurate fusion identification.
- Practical guidance on assay design, QC, and validation supports routine clinical integration.
