Background/Objectives: Mesenchymal–epithelial transition (MET) gene amplification is a critical biomarker in non-small cell lung cancer (NSCLC), significantly influencing treatment decisions and prognostic evaluations. However, current detection methods such as fluorescence in situ hybridization (FISH) and next-generation sequencing (NGS) have limitations in speed, cost, and specificity, particularly when distinguishing between focalMETamplification andMETpolysomy. Methods: This study introduces a novel digital PCR (dPCR) assay designed not only to detectMETamplification but also to differentiate between its focal and non-focal subtypes. The assay was evaluated against established FISH and targeted NGS panels using 55 NSCLC samples with knownMETamplification statuses (26 positive and 29 negative) confirmed by FISH and NGS. Results The dPCR assay demonstrated high sensitivity (96.0%) and specificity (96.7%), achieving 100% concordance with FISH in differentiating focalMETamplification fromMETpolysomy. Additionally, the assay exhibited excellent precision, accuracy, and linearity (R2= 1.00) inMETcopy number quantification, surpassing NGS in diagnostic performance. Offering a robust, cost-effective, and efficient alternative to FISH, the dPCR assay significantly reduces the turnaround time (3 h versus 2 days) and provides a quantitative and objective method forMETamplification detection and subtype differentiation. This makes it suitable for clinical laboratories with limited molecular expertise. Conclusions: This study highlights the potential of the dPCR assay to complement existing molecular diagnostic techniques, delivering reliable and actionable results forMET-targeted therapy selection in NSCLC patients and thereby advancing precision oncology.
背景/目的：间质上皮转化（MET）基因扩增是非小细胞肺癌（NSCLC）的关键生物标志物，显著影响治疗决策和预后评估。然而，目前荧光原位杂交（FISH）和二代测序（NGS）等检测方法在速度、成本和特异性方面存在局限，尤其在区分局灶性MET扩增与MET多体性时。方法：本研究开发了一种新型数字PCR（dPCR）检测方法，不仅能检测MET扩增，还能区分其局灶性和非局灶性亚型。该方法通过55例经FISH和NGS确认MET扩增状态（26例阳性，29例阴性）的NSCLC样本，与已建立的FISH和靶向NGS检测板进行了对比评估。结果：dPCR检测显示出高灵敏度（96.0%）和高特异性（96.7%），在区分局灶性MET扩增与MET多体性方面与FISH结果完全一致（100%）。此外，该方法在MET拷贝数定量方面表现出优异的精密度、准确性和线性（R²=1.00），诊断性能优于NGS。作为FISH的稳健、经济高效且快速的替代方案，dPCR检测显著缩短了检测周期（3小时对比2天），并为MET扩增检测和亚型区分提供了定量、客观的方法，适用于分子专业知识有限的临床实验室。结论：本研究强调了dPCR检测作为现有分子诊断技术补充的潜力，能为NSCLC患者MET靶向治疗选择提供可靠且可操作的检测结果，从而推动精准肿瘤学的发展。
肿瘤(癌症)患者之家