title: A new rapid method for detecting epidermal growth factor receptor mutations in non-small cell lung cancer.
abstract: Mutations in the epidermal growth factor receptor (EGFR) gene are associated with a favorable clinical response to the EGFR tyrosine kinase inhibitors gefitinib and erlotinib in non-small cell lung cancer (NSCLC). We present here, a new method for the rapid detection of the two most common EGFR mutations (delE746-A750 and L858R) from clinical samples. The methodology involves the combination of newly designed mutation-specific primers and a novel real-time PCR machine with an innovative thermo-control mechanism that enables ultrarapid PCR. We evaluated this method using a cell mixture composed of various ratios of lung cancer cells harboring mutated or wild-type EGFR, lung cancer tissues obtained by surgery, and a cytology sample obtained by bronchoscopy from a lung cancer patient. In the cell mixture analysis, our method detected 0.1% of cells with delE746-A750 and 1% of cells with L858R among cells with wild-type EGFR. In 143 lung cancer tissues, the result of this assay was concordant with those of direct sequencing in 138 samples. The five samples with discordant results were tested using a PCR-Invader assay and the result matched those of our method at 100%. We also successfully detected EGFR mutations in the lavage obtained from a lung cancer patient. The turnaround time for this method was <10 min, and all steps could be accomplished in <50 min after sample collection. Thus, our novel PCR method offers a rapid, simple, and less expensive test for EGFR mutations and can be applied as a point-of-care diagnostic test.
authors.affiliation: Division of Medical Oncology and Molecular Respirology, Department of Multidisciplinary Internal Medicine, Tottori University, Yonago-shi, Tottori-ken, Japan.
authors.forename: Miyako
authors.initials: M
authors.lastname: Takata
authors.affiliation: Division of Medical Oncology and Molecular Respirology, Department of Multidisciplinary Internal Medicine, Tottori University, Yonago-shi, Tottori-ken, Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Chikumi
authors.affiliation: Division of Medical Oncology and Molecular Respirology, Department of Multidisciplinary Internal Medicine, Tottori University, Yonago-shi, Tottori-ken, Japan.
authors.forename: Keiji
authors.initials: K
authors.lastname: Matsunami
authors.affiliation: Division of Medical Oncology and Molecular Respirology, Department of Multidisciplinary Internal Medicine, Tottori University, Yonago-shi, Tottori-ken, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Kodani
authors.affiliation: Division of Medical Oncology and Molecular Respirology, Department of Multidisciplinary Internal Medicine, Tottori University, Yonago-shi, Tottori-ken, Japan.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Sakamoto
authors.affiliation: Trust Medical Co., Ltd., Hyogo, Japan.
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Hashimoto
authors.affiliation: Division of Medical Oncology and Molecular Respirology, Department of Multidisciplinary Internal Medicine, Tottori University, Yonago-shi, Tottori-ken, Japan.
authors.forename: Masaki
authors.initials: M
authors.lastname: Nakamoto
authors.affiliation: Division of Medical Oncology and Molecular Respirology, Department of Multidisciplinary Internal Medicine, Tottori University, Yonago-shi, Tottori-ken, Japan.
authors.forename: Kensaku
authors.initials: K
authors.lastname: Okada
authors.affiliation: Division of Medical Oncology and Molecular Respirology, Department of Multidisciplinary Internal Medicine, Tottori University, Yonago-shi, Tottori-ken, Japan.
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Kitaura
authors.affiliation: Exploratory Oncology Research and Clinical Trial Center, National Cancer Center, Kashiwa, Chiba, Japan.
authors.forename: Shingo
authors.initials: S
authors.lastname: Matsumoto
authors.affiliation: Division of Medical Oncology and Molecular Respirology, Department of Multidisciplinary Internal Medicine, Tottori University, Yonago-shi, Tottori-ken, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Kurai
authors.affiliation: Division of Medical Oncology and Molecular Respirology, Department of Multidisciplinary Internal Medicine, Tottori University, Yonago-shi, Tottori-ken, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Yamasaki
authors.affiliation: Division of Medical Oncology and Molecular Respirology, Department of Multidisciplinary Internal Medicine, Tottori University, Yonago-shi, Tottori-ken, Japan.
authors.forename: Tadashi
authors.initials: T
authors.lastname: Igishi
authors.affiliation: Division of Medical Oncology and Molecular Respirology, Department of Multidisciplinary Internal Medicine, Tottori University, Yonago-shi, Tottori-ken, Japan.
authors.forename: Naoto
authors.initials: N
authors.lastname: Burioka
authors.affiliation: Division of Medical Oncology and Molecular Respirology, Department of Multidisciplinary Internal Medicine, Tottori University, Yonago-shi, Tottori-ken, Japan.
authors.forename: Eiji
authors.initials: E
authors.lastname: Shimizu
mesh_terms: D000328:Adult; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods*; D066246:ErbB Receptors / Q000235:genetics*; D005260:Female; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics*; D009154:Mutation; D019095:Point-of-Care Systems; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*
chemical_list: C512478:EGFR protein, human; D066246:ErbB Receptors