J Cancer 2019; 10(18):4341-4349. doi:10.7150/jca.31326

Research Paper

Detection of Plasma EGFR Mutations in NSCLC Patients with a Validated ddPCR Lung cfDNA Assay

Qiao-mei Guo1#, Lin Wang1#, Wen-jun Yu1, Li-hua Qiao1, Ming-na Zhao1, Xiao-meng Hu1, Ya-meng Sun3, Sheng Ni3, Yun-hua Xu2✉, Jia-tao Lou1✉

1. Department of Laboratory Medicine, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai, China
2. Shanghai Lung Cancer Center, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai, China
3. Bio-Chain Biological Technology Co., Ltd, Shanghai, China
#Qiao-mei Guo and Lin Wang are the first co-authors and they contributed equally to this work.

Abstract

Purpose: The clinical utility of cell-free DNA (cfDNA) to assess EGFR mutations is increasing. However, there are limited studies determining their clinical validity and utility. The value of cfDNA assays in cancer management remains controversial.

Methods: In this study, we first evaluated the analytical performance of the ddPCR Lung cfDNA Assay. We next analyzed the concordance of the results with tissue amplification refractory mutation system PCR (ARMS-PCR) and plasma next-generation sequencing (NGS) genotyping. Finally, we assessed its clinical utility by exploring the association of cfDNA EGFR mutations with metastatic sites and the efficacy of EGFR-TKIs treatment.

Results: The ddPCR Lung cfDNA Assay demonstrated a limit of blank of 1 droplet per reaction, an analytical specificity of 100%, and detection limit of 0.05%, 0.05%, and 0.1% for E746_A750del, L858R, and T790M, respectively. With tissue ARMS-PCR as a standard for comparison, the clinical sensitivity and specificity of ddPCR were 62.5% (15/24) and 100% (82/82) for E746_A750del, and 75.0% (15/20) and 94.2% (81/86) for L858R, respectively. The ddPCR showed high concordance with NGS in determining cfDNA EGFR mutations. Patients with bone and/or brain metastasis showed a higher detection rate and mutant abundance of cfDNA EGFR mutations compared to those with other sites of metastasis. Moreover, EGFR-TKIs treatment was effective in patients with sensitive EGFR mutations in either plasma cfDNA or tumor tissue-derived DNA.

Conclusions: We validated in this study that the ddPCR Lung cfDNA Assay is reliable for detection of EGFR mutations in lung cancers, in terms of analytical performance, clinical validity and utility.

Keywords: cell-free DNA, droplet digital PCR, EGFR, NGS, non-small cell lung cancer

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How to cite this article:
Guo Qm, Wang L, Yu Wj, Qiao Lh, Zhao Mn, Hu Xm, Sun Ym, Ni S, Xu Yh, Lou Jt. Detection of Plasma EGFR Mutations in NSCLC Patients with a Validated ddPCR Lung cfDNA Assay. J Cancer 2019; 10(18):4341-4349. doi:10.7150/jca.31326. Available from http://www.jcancer.org/v10p4341.htm