Real-world performance of next-generation sequencing in advanced non-squamous non-small cell lung cancer in Brazil.

BACKGROUND

Broad molecular profiling via next-generation sequencing (NGS) is essential for guiding targeted therapy in advanced non-squamous non-small cell lung cancer (nsNSCLC). However, real-world implementation in low- and middle-income countries remains limited by operational challenges.

METHODS

We retrospectively analyzed data on nsNSCLC samples tested at a centralized Brazilian genomics laboratory between 2021 and 2025, evaluating NGS performance over time, causes of failure, and the impact of workflow improvements.

RESULTS

A total of 2522 nsNSCLC samples were analyzed between 2021 and 2025. Overall NGS success increased from 64.2% to 85.7% (p < 0.0001) following workflow optimization and the adoption of a hybrid-capture platform with lower DNA input requirement. Insufficient tissue and suboptimal nucleic acid quality were the main causes of failure throughout the study period. Failure rates decreased across all Brazilian regions, although referring laboratories of low volume continued to underperform due to persistent pre-analytical factors.

CONCLUSION

Laboratory workflow optimization, particularly the adoption of hybrid-capture methodology, significantly improved NGS success rates in a nationwide nsNSCLC testing program. Persistent high failure rates in low-volume centers underscores the need for targeted education and infrastructural investment to broaden equitable access to precision oncology in Brazil.