Picoliter-Droplet Digital Polymerase Chain Reaction-Based Analysis of Cell-Free Plasma DNA to Assess EGFR Mutations in Lung Adenocarcinoma That Confer Resistance to Tyrosine-Kinase Inhibitors

摘要: PURPOSE The objective of this study was to evaluate the utility analyzing cell-free plasma DNA (cfDNA) by picoliter-droplet digital polymerase chain reaction (ddPCR) detect EGFR mutations that confer resistance tyrosine-kinase inhibitors (TKIs) used for treatment lung adenocarcinoma (LADC). EXPERIMENTAL DESIGN Thirty-five LADC patients who received epidermal growth factor receptor (EGFR)-TKI therapy, including ten tumor rebiopsy after development resistance, were subjected picoliter-ddPCR-cfDNA analysis determine fraction cfDNA with TKI-sensitive (L858R and inflame exon 19 deletions) -resistant (i.e., T790M) mutations, as well their concordance mutation status in rebiopsied tissues. RESULTS samples from 15 (94%) 16 acquired positive mutations. Also, 7 (44%) T790M mutation, fractions (+) ranging 7.4% 97%. positivity consistent eight whom tissues analyzed, whereas remaining cases negative tumors. Prior EGFR-TKI cfDNAs 9 (38%) 0 24 respectively. Next-generation sequencing one patient exhibited innate TKI despite a high absence his revealed presence L747P known driver resistance. CONCLUSION Picoliter-ddPCR examination cfDNA, supported next-generation analysis, enables noninvasive assessment TKIs. IMPLICATIONS FOR PRACTICE Noninvasive monitoring predominance tumors harboring secondary activating gene is necessary precise effective adenocarcinoma. Because cells are resistant receptor-tyrosine-kinase (TKIs), influences choice whether use conventional or Digital reaction-based promising method; however, its feasibility, consistency tissue, has not been fully proven. Here, technology presented candidate method testing assessing T790M-mutant