Mirikar Duhita, Banerjee Nandini, Prabhash Kumar, Kaushal Rajiv Kumar, Naronha Vanita, Pramesh C S, Karimundackal George, Joshi Amit, Rane Swapnil, Basak Ranjan
Translational Research Laboratory, Advanced Centre for Treatment, Research & Education in Cancer, Tata Memorial Centre, Navi Mumbai, India.
University of North Carolina, Charlotte, USA.
Sci Rep. 2025 Jan 8;15(1):1252. doi: 10.1038/s41598-025-85160-6.
In this study, we measured human epidermal growth factor receptor (EGFR) mutations in both tissue and circulating tumor DNA (ctDNA) by using beads, emulsions, amplifications and magnetic polymerase chain reaction (BEAMing PCR). Noninvasive mutation detection by assessing circulating tumor DNA (ctDNA) offers many advantages over tumor biopsy. One hundred non-small cell lung cancer (NSCLC) patients were enrolled, and both preoperative plasma samples and formalin-fixed and paraffin-embedded (FFPE) samples were collected for the study. The EGFR mutation status was determined by BEAMing PCR in ctDNA. Real-time quantitative PCR (qPCR) data were collected from our hospital database (EMR-qPCR, Electronic Medical Records) for comparative analysis. Additionally, qPCR was also performed on FFPE tissues using a Diatech EGFR qPCR kit. The concordance rates were 98.8%, 98.9% and 95.5% for exons 19, 20 and 21, respectively, when the BEAMing data were compared with the EMR-qPCR data. Additionally, when the BEAMing and Diatech qPCR data were compared, 90%, 100%, 96% and 98% of the genes were obtained for exons 19, 20, 21 (L858R) and 21 (L861Q), respectively. For both comparisons, Cohen's kappa agreement was significant. The advantage of BEAMing is its ability to identify mutated DNA sequences in cancer cells in the background of normal cell DNA contamination. This could be useful for disease monitoring and progression.
在本研究中，我们使用磁珠、乳液、扩增和磁性聚合酶链反应（BEAMing PCR）技术检测了组织和循环肿瘤DNA（ctDNA）中的人类表皮生长因子受体（EGFR）突变。通过评估循环肿瘤DNA（ctDNA）进行无创突变检测比肿瘤活检具有许多优势。招募了100例非小细胞肺癌（NSCLC）患者，收集术前血浆样本以及福尔马林固定石蜡包埋（FFPE）样本用于研究。通过BEAMing PCR确定ctDNA中的EGFR突变状态。从我们医院数据库（电子病历-qPCR，EMR-qPCR）收集实时定量PCR（qPCR）数据用于比较分析。此外，还使用Diatech EGFR qPCR试剂盒对FFPE组织进行qPCR。当将BEAMing数据与EMR-qPCR数据进行比较时，外显子19、20和21的一致性率分别为98.8%、98.9%和95.5%。此外，当比较BEAMing和Diatech qPCR数据时，外显子19、20、21（L858R）和21（L861Q）分别获得了90%、100%、96%和98%的基因一致性。对于这两种比较，科恩kappa一致性均具有显著性。BEAMing的优势在于其能够在正常细胞DNA污染背景下识别癌细胞中的突变DNA序列。这对于疾病监测和病情进展可能有用。
