Molecular Pathology Division, Department of Pathology, Tata Memorial Hospital, Homi Bhabha National Institute, Mumbai, India.
Department of Pathology, Tata Memorial Hospital, Homi Bhabha National Institute, Mumbai, India.
Int J Exp Pathol. 2024 Jun;105(3):90-99. doi: 10.1111/iep.12503. Epub 2024 May 8.
Management of lung cancer today obligates a mutational analysis of the epidermal growth factor receptor (EGFR) gene particularly when Tyrosine Kinase Inhibitor (TKI) therapy is being considered as part of prognostic stratification. This study evaluates the performance of automated microfluidics-based EGFR mutation detection and its significance in clinical diagnostic settings. Formalin-fixed, paraffin-embedded (FFPE) samples from NSCLC patients (n = 174) were included in a two-phase study. Phase I: Validation of the platform by comparing the results with conventional real-time PCR and next-generation sequencing (NGS) platform. Phase II: EGFR mutation detection on microfluidics-based platform as part of routine diagnostics workup. The microfluidics-based platform demonstrates 96.5% and 89.2% concordance with conventional real-time PCR and NGS, respectively. The system efficiently detects mutations across the EGFR gene with 88.23% sensitivity and 100% specificity. Out of 144 samples analysed in phase II, the platform generated valid results in 94% with mutation detected in 41% of samples. This microfluidics-based platform can detect as low as 5% mutant allele fractions from the FFPE samples. Therefore the microfluidics-based platform is a rapid, complete walkaway, with minimum tissue requirement (two sections of 5 μ thickness) and technical skill requirement. The method can detect clinically actionable EGFR mutations efficiently and can be considered a reliable diagnostic platform in resource-limited settings. From receiving samples to reporting the results this platform provides accurate data without much manual intervention. The study helped to devise an algorithm that emphasizes effective screening of the NSCLC cases for EGFR mutations with varying tumour content. Thus it helps in triaging the cases judiciously before proceeding with multigene testing.
如今，在考虑将酪氨酸激酶抑制剂（TKI）治疗作为预后分层的一部分时，肺癌的管理需要对表皮生长因子受体（EGFR）基因进行突变分析。本研究评估了自动化微流控 EGFR 突变检测的性能及其在临床诊断环境中的意义。本研究纳入了 174 名非小细胞肺癌（NSCLC）患者的福尔马林固定、石蜡包埋（FFPE）样本，分两个阶段进行：第一阶段：通过与传统实时 PCR 和下一代测序（NGS）平台的结果进行比较来验证平台；第二阶段：在微流控平台上进行 EGFR 突变检测，作为常规诊断工作的一部分。微流控平台与传统实时 PCR 和 NGS 的一致性分别为 96.5%和 89.2%。该系统能够有效地检测 EGFR 基因中的突变，灵敏度为 88.23%，特异性为 100%。在第二阶段分析的 144 个样本中，该平台在 94%的样本中产生了有效的结果，其中 41%的样本检测到了突变。该微流控平台可以从 FFPE 样本中检测到低至 5%的突变等位基因分数。因此，该微流控平台是一种快速、完整的自动化平台，对组织的需求最小（两个 5μm 厚的切片），技术要求也最低。该方法能够有效地检测出临床可操作的 EGFR 突变，可作为资源有限环境下可靠的诊断平台。从接收样本到报告结果，该平台提供了准确的数据，几乎不需要人工干预。该研究有助于制定一种算法，强调对 EGFR 突变的 NSCLC 病例进行有效筛选，根据肿瘤含量的不同对病例进行分类。因此，在进行多基因检测之前，有助于明智地对病例进行分诊。
