Developing Ultrasensitive Library-Aliquot-Based Droplet Digital PCR for Detecting T790M in Plasma-Circulating Tumor DNA of Non-small-Cell-Lung-Cancer Patients

Ke Yang, Jun Li, Jiuzhou Zhao, Pengfei Ren, Zhizhong Wang, Bing Wei, Bing Dong, Rui Sun, Xiaoyan Wang, Harry J M Groen, Jie Ma, Yongjun Guo

Research output: Contribution to journalArticleAcademicpeer-review

10 Citations (Scopus)

Abstract

A T790M secondary mutation in epidermalgrowth-factor receptor (EGFR) is the most well-established EGFR-tyrosine-kinase-inhibitor (TKI) resistance marker in non-small-cell lung cancer (NSCLC). The current methods to rapidly and accurately detect T790M in clinical practice are not satisfactory because of several obstacles, including the unavailability of tumor-tissue rebiopsies and the low DNA copy number of T790M in circulating tumor DNA (ctDNA). Here, we develop library-aliquot-based droplet digital PCR (LAB-ddPCR) to increase detection sensitivity without affecting accuracy. This new LAB-ddPCR method is performed using aliquots of the ctDNA precapture nextgeneration-sequencing (NGS) library, in which the isolated ctDNA was amplified and enriched. We show that the LAB-ddPCR can precisely distinguish between T790M wild-type and mutation alleles without introducing extra false-positive signals. In a cohort of 70 post-TKI NSCLC patients, the LAB-ddPCR identified 41 T790M-positive cases (sensitivity 58.57%), but ddPCR only detected T790M in 27 cases (sensitivity 38.57%). Taking the ARMS-PCR result from matched tumor rebiopsies into consideration, the LAB-ddPCR method is better than ddPCR. In conclusion, the LAB-ddPCR ctDNA test offers a feasible and flexible option for the rapid and accurate detection of the T790M secondary mutation, which is helpful in dynamically monitoring drug response and disease progression throughout the therapeutic regimen.

Original languageEnglish
Pages (from-to)11203-11209
Number of pages7
JournalAnalytical Chemistry
Volume90
Issue number19
DOIs
Publication statusPublished - 2-Oct-2018

Keywords

  • EGFR T790M
  • CTDNA
  • RESISTANCE
  • MUTATION
  • TKI
  • GEFITINIB
  • ERLOTINIB
  • STANDARD
  • OUTCOMES
  • AZD9291

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