title: Application of Single-Molecule Amplification and Resequencing Technology for Broad Surveillance of Plasma Mutations in Patients with Advanced Lung Adenocarcinoma.
abstract: Liquid biopsy to access the circulating tumor DNA is a promising surrogate for invasive tumor genotyping. We designed a multiplex assay based on circulating single-molecule amplification and resequencing technology (cSMART) to simultaneously detect and quantitate hot spot EGFR, KRAS, BRAF, ERBB2, and ALK plasma DNA variants in 103 patients with advanced lung adenocarcinoma. In validation studies using an analytical mutation standard, the sensitivity of the assay for EGFR mutation detection was at least 0.1% and specificity was 100%. The diagnostic detection sensitivity was one mutant molecule per 2 mL of plasma. The most frequently detected plasma mutations were EGFR variants L858R (21.4%), exon 19 deletions (19.4%), T790M (9.7%), and KRAS G12X variants (9.7%). Rarer were BRAF V600X (1.95%) and ERBB2 exon 20 (0.97%) variants. In single samples, four novel EGFR exon 19 deletions, one KIF5B-ALK, and two EML4-ALK variants were also detected. From comparisons of 103 matched plasma and tumor specimen genotypes, 75 (72.8%) were concordant, 9 (8.8%) were partially concordant, and 19 (18.4%) were discordant. Overall, the combined positive and negative concordance rate for detection of each oncogenic variant exceeded 90%. On the basis of these findings, we propose that cSMART displays the diagnostic hallmarks of a comprehensive plasma genotyping assay, with potential application for precisely monitoring changes in plasma mutation levels in response to targeted drug therapy.
authors.affiliation: Department of Pathology, Beijing Hospital, National Center of Gerontology, Beijing, China.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Department of Medical Oncology, Beijing Hospital, National Center of Gerontology, Beijing, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Cheng
authors.affiliation: Department of Medical Oncology, Cancer Institute and Hospital Chinese Academy of Medical Sciences, Beijing, China.
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Han
authors.affiliation: Department of Respiratory and Critical Care Medicine, Peking University People's Hospital, Beijing, China.
authors.forename: Xinlin
authors.initials: X
authors.lastname: Mu
authors.affiliation: Beijing Institute of Respiratory Medicine, Beijing Chao-Yang Hospital, Capital Medical University, Beijing, China.
authors.forename: Yuhui
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Pathology, Beijing Hospital, National Center of Gerontology, Beijing, China.
authors.forename: Di
authors.initials: D
authors.lastname: Cui
authors.affiliation: Department of Medical Oncology, Military General Hospital of Beijing PLA, Beijing, China.
authors.forename: Chang
authors.initials: C
authors.lastname: Liu
authors.affiliation: Department of Respiratory Medicine, Peking Union Medical College Hospital, Beijing, China.
authors.forename: Li
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Medical Oncology, Air Force General Hospital of Beijing PLA, Beijing, China.
authors.forename: Zaiwen
authors.initials: Z
authors.lastname: Fan
authors.affiliation: Department of Respiratory Medicine, The First Affiliated Hospital of General Hospital of PLA, Beijing, China.
authors.forename: Lingyun
authors.initials: L
authors.lastname: Ma
authors.affiliation: Department of Pathology, Beijing Hospital, National Center of Gerontology, Beijing, China.
authors.forename: Li
authors.initials: L
authors.lastname: Yang
authors.affiliation: Department of Pathology, Beijing Hospital, National Center of Gerontology, Beijing, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Di
authors.affiliation: Research and Development Department, Berry Genomics Corp., Beijing, China.
authors.forename: David S
authors.initials: DS
authors.lastname: Cram
authors.affiliation: Department of Medical Oncology, Cancer Institute and Hospital Chinese Academy of Medical Sciences, Beijing, China. Electronic address: syuankaipumc@126.com.
authors.forename: Yuankai
authors.initials: Y
authors.lastname: Shi
authors.affiliation: Department of Pathology, Beijing Hospital, National Center of Gerontology, Beijing, China. Electronic address: liudongge@sohu.com.
authors.forename: Dongge
authors.initials: D
authors.lastname: Liu
mesh_terms: D000230:Adenocarcinoma / Q000097:blood / Q000175:diagnosis / Q000235:genetics*; D000077192:Adenocarcinoma of Lung; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001483:Base Sequence; D016022:Case-Control Studies; D004252:DNA Mutational Analysis; D004273:DNA, Neoplasm / Q000097:blood / Q000235:genetics* / Q000302:isolation & purification; D005260:Female; D052138:Genes, Neoplasm; D006801:Humans; D008175:Lung Neoplasms / Q000097:blood / Q000175:diagnosis / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques; D009154:Mutation; D009367:Neoplasm Staging; D012680:Sensitivity and Specificity; D055815:Young Adult
chemical_list: D004273:DNA, Neoplasm