title: EGFR Mutation Analysis of Circulating Tumor DNA Using an Improved PNA-LNA PCR Clamp Method.
abstract: Introduction. Rebiopsies have become more crucial in non-small cell lung cancer (NSCLC). Instead of invasive biopsies, development of collecting biological data of the tumor from blood samples is expected. We conducted a prospective study to assess the feasibility of detection of epidermal growth factor receptor (EGFR) mutation in plasma samples. Method. NSCLC patients harboring EGFR activating mutations, who were going to receive EGFR-tyrosine kinase inhibitors (TKIs) as first-line treatment, were enrolled in this study. Plasma EGFR activating mutations and the T790M resistance mutation were analyzed by an improved PNA-LNA PCR clamp method, characterized by a 10-fold or more sensitivity compared with the original methods. Result. Six patients with wild-type EGFR and 24 patients with EGFR mutations were enrolled in this study. Pretreatment plasma samples achieved sensitivity of 79%. The 6 patients with wild-type EGFR were all negative for plasma EGFR mutations. At the time of disease progression, plasma T790M mutation was detected in 8 of 16 cases. Absence of T790M before and during TKI treatment and disappearance of activating mutations during TKI treatment were considered as predictors of EGFR-TKIs efficacy. Conclusion. We were able to detect EGFR mutations in plasma samples by using an improved PNA-LNA PCR clamp method.
authors.affiliation: Department of Respiratory Medicine, Miyagi Cancer Center, 47-1 Nodayama, Medeshima-Shiote, Natori 981-1293, Japan.
authors.forename: Kana
authors.initials: K
authors.lastname: Watanabe
authors.affiliation: Department of Respiratory Medicine, Miyagi Cancer Center, 47-1 Nodayama, Medeshima-Shiote, Natori 981-1293, Japan.
authors.forename: Tatsuro
authors.initials: T
authors.lastname: Fukuhara
authors.affiliation: Department of Respiratory Medicine, Miyagi Cancer Center, 47-1 Nodayama, Medeshima-Shiote, Natori 981-1293, Japan.
authors.forename: Yoko
authors.initials: Y
authors.lastname: Tsukita
authors.affiliation: Department of Respiratory Medicine, Miyagi Cancer Center, 47-1 Nodayama, Medeshima-Shiote, Natori 981-1293, Japan.
authors.forename: Mami
authors.initials: M
authors.lastname: Morita
authors.affiliation: Department of Respiratory Medicine, Miyagi Cancer Center, 47-1 Nodayama, Medeshima-Shiote, Natori 981-1293, Japan.
authors.forename: Aya
authors.initials: A
authors.lastname: Suzuki
authors.affiliation: Division of Cancer Biology and Therapeutics, Miyagi Cancer Center Research Institute, Natori, Japan.
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Tanaka
authors.affiliation: Molecular Genetic Research Department, LSI Medience Corporation, Tokyo, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Terasaki
authors.affiliation: Japan Anti-Tuberculosis Association, Tokyo, Japan.
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Nukiwa
authors.affiliation: Department of Respiratory Medicine, Miyagi Cancer Center, 47-1 Nodayama, Medeshima-Shiote, Natori 981-1293, Japan.
authors.forename: Makoto
authors.identifier: 0000-0002-1950-1943
authors.initials: M
authors.lastname: Maemondo
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D000970:Antineoplastic Agents / Q000494:pharmacology / Q000627:therapeutic use; D002289:Carcinoma, Non-Small-Cell Lung / Q000188:drug therapy / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods*; D004273:DNA, Neoplasm / Q000097:blood*; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D066246:ErbB Receptors / Q000037:antagonists & inhibitors; D005260:Female; D000077156:Gefitinib; D018773:Genes, erbB-1; D006801:Humans; D008175:Lung Neoplasms / Q000188:drug therapy / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D011446:Prospective Studies; D011799:Quinazolines / Q000494:pharmacology / Q000627:therapeutic use
chemical_list: D000970:Antineoplastic Agents; D004273:DNA, Neoplasm; D011799:Quinazolines; D066246:ErbB Receptors; D000077156:Gefitinib