Somekawa Kohei, Kobayashi Nobuaki, Nagaoka Satoshi, Seki Kenichi, Kajita Yukihito, Muraoka Suguru, Izawa Ami, Kaneko Ayami, Otsu Yukiko, Hirata Momo, Kubo Sousuke, Nagasawa Ryo, Murohashi Kota, Fuji Hiroaki, Teranishi Shuhei, Tashiro Ken, Watanabe Keisuke, Horita Nobuyuki, Hara Yu, Kudo Makoto, Kaneko Takeshi
Department of Pulmonology, Yokohama City University Graduate School of Medicine, Yokohama, Japan.
Respiratory Disease Center, Yokohama City University Medical Center, Yokohama, Japan.
Transl Lung Cancer Res. 2025 Feb 28;14(2):353-362. doi: 10.21037/tlcr-24-772. Epub 2025 Feb 22.
Molecular profiling of non-small cell lung cancer (NSCLC) is crucial for personalized treatment, but obtaining adequate tumor tissue can be challenging. This study evaluated the utility of droplet digital polymerase chain reaction (ddPCR) analysis of bronchial washings (BWs) and serum for detecting driver oncogene mutations in NSCLC patients, comparing its performance to standard tissue genotyping methods.
In this prospective, multicenter study conducted at two university hospitals in Yokohama, Japan, 73 treatment-naïve NSCLC patients underwent bronchoscopy with BW collection and blood sampling between October 2022 and April 2024. ddPCR was performed on BW and serum samples to detect epidermal growth factor receptor (EGFR; L858R, exon 19 deletions, G719X), KRAS (G12/13), and BRAF (V600E) mutations. Results were compared with standard tissue genotyping methods, including AmoyDx and Oncomine Dx Target Test (DxTT) assays. Turnaround time (TAT) for results was also assessed. The study protocol was approved by the institutional review boards, and all participants provided informed consent.
ddPCR analysis of BW samples showed high concordance with tissue genotyping, detecting EGFR mutations in 31.5% of cases (identical to tissue). For common EGFR mutations (L858R and exon 19 deletions), BW genotyping demonstrated 100% sensitivity and 98.0% specificity compared to tissue. TAT was significantly shorter for BW ddPCR compared to tissue genotyping (4.4±1.8 20.4±7.7 days, P<0.001). Serum ddPCR showed lower sensitivity (7.8% 33.3% for EGFR mutations) compared to tissue genotyping, with detection associated with the presence of bone metastases. KRAS and BRAF mutations were detected at similar rates in BW and tissue samples, but at lower rates in serum.
ddPCR analysis of BWs demonstrates high accuracy and rapid TAT for detecting common driver mutations in NSCLC. This approach represents a promising alternative to tissue biopsy for molecular profiling, potentially expediting treatment decisions. While serum ddPCR showed limited utility, it may complement tissue genotyping in specific clinical scenarios.
非小细胞肺癌（NSCLC）的分子谱分析对于个性化治疗至关重要，但获取足够的肿瘤组织可能具有挑战性。本研究评估了支气管灌洗（BW）和血清的液滴数字聚合酶链反应（ddPCR）分析在检测NSCLC患者驱动癌基因突变方面的效用，并将其性能与标准组织基因分型方法进行比较。
在日本横滨的两家大学医院进行的这项前瞻性多中心研究中，73例未经治疗的NSCLC患者在2022年10月至2024年4月期间接受了支气管镜检查，同时采集了BW样本和血液样本。对BW和血清样本进行ddPCR检测，以检测表皮生长因子受体（EGFR；L858R、外显子19缺失、G719X）、KRAS（G12/13）和BRAF（V600E）突变。将结果与标准组织基因分型方法进行比较，包括厦门艾德和安可美达DxTT检测。还评估了结果的周转时间（TAT）。该研究方案已获得机构审查委员会的批准，所有参与者均提供了知情同意书。
BW样本的ddPCR分析与组织基因分型显示出高度一致性，在31.5%的病例中检测到EGFR突变（与组织相同）。对于常见的EGFR突变（L858R和外显子19缺失），与组织相比，BW基因分型显示出100%的敏感性和98.0%的特异性。与组织基因分型相比，BW的ddPCR的TAT明显更短（4.4±1.8对20.4±7.7天，P<0.001）。与组织基因分型相比，血清ddPCR的敏感性较低（EGFR突变的敏感性为7.8%对33.3%），检测与骨转移的存在相关。BW和组织样本中KRAS和BRAF突变的检测率相似，但血清中的检测率较低。
BW的ddPCR分析在检测NSCLC常见驱动突变方面显示出高准确性和快速的TAT。这种方法是分子谱分析组织活检的一种有前途的替代方法，可能会加快治疗决策。虽然血清ddPCR的效用有限，但在特定临床情况下可能会补充组织基因分型。
