Journal Article

Suitability of Frozen Pleural Fluid Pellets for Next‐Generation Sequencing‐Based Driver Gene Testing in Non‐Small Cell Lung Cancer

Iwama, Eiji, Okamoto, Isamu, Nakashima, Kazuki, Tsuchiya‐Kawano, Yuko, Otsubo, Kohei, Harada, Eiji, Mikumo, Hironori

2025

Overview

Background Driver gene alterations are increasingly being identified, and multiplex genetic tests have become essential for determining the optimal treatment method for advanced non‐small cell lung cancer (NSCLC). Next‐generation sequencing (NGS) enables the simultaneous detection of multiple driver gene alterations using nucleic acids extracted from tumor tissue samples. However, obtaining sufficient tumor tissue volume is challenging, and inadequate formalin fixation can lead to the failure of NGS analysis. Malignant pleural effusions can be collected using a simple, minimally invasive technique; however, it remains uncertain whether pleural fluid is a suitable source for detecting driver gene alterations using NGS. This retrospective observational study examined the suitability of fresh‐frozen pleural fluid pellets for NGS in clinical practice. Methods Patients with NSCLC whose frozen pleural fluid pellets were analyzed using the Oncomine Dx Target Test Multi‐CDx System between June 2019 and February 2024 were included in the study. The primary endpoint was the success rate of driver gene analysis. Results In total, 26 patients were enrolled. The success rate for testing alterations in driver genes was 92.3% (24/26), and the detection rate of targetable driver gene alterations was 53.8% (14/26). The median turnaround time from sample submission to result confirmation was 10 days (range 7–19 days). Conclusion Our findings indicate that frozen pleural fluid pellets are suitable for NGS‐based driver gene testing in patients with advanced NSCLC. This approach provides a practical alternative for multigene testing when sufficient tissue is not available. This study demonstrated a high success rate of driver gene testing using the Oncomine Dx Target Test (ODxTT) on fresh‐frozen pleural fluid pellets from patients with NSCLC.

• Publisher: John Wiley & Sons Australia, Ltd,John Wiley & Sons, Inc,Wiley
• Subject: Adult / Aged / Aged, 80 and over / Analysis / Antimitotic agents / Antineoplastic agents / Biopsy / Cancer / Carcinoma, Non-Small-Cell Lung - genetics / Carcinoma, Non-Small-Cell Lung - pathology / Cellular biology / driver gene alterations / Female / frozen pleural fluid pellet / Genes / Genetic aspects / Genetic screening / Genetic testing / Genetic Testing - methods / High-Throughput Nucleotide Sequencing - methods / Histology / Humans / Kinases / Lung cancer / Lung cancer, Non-small cell / Lung cancer, Small cell / Lung Neoplasms - genetics / Lung Neoplasms - pathology / Male / Middle Aged / Mutation / next‐generation sequencing / non‐small cell lung cancer / Nucleic acids / Original / Patients / Pleural effusion / Pleural Effusion, Malignant - genetics / Pleural Effusion, Malignant - pathology / Retrospective Studies / Success / tumor sample