title: Detection of EGFR mutations in archived cytologic specimens of non-small cell lung cancer using high-resolution melting analysis.
abstract: Mutations of the epidermal growth factor receptor (EGFR), particularly deletional mutations (DEL) in exon 19 and L858R in exon 21, are reportedly correlated with clinical outcome in patients with non-small cell lung cancer (NSCLC) receiving the EGFR tyrosine kinase inhibitors gefitinib and erlotinib, suggesting that detection of EGFR mutations would have an important role in clinical decision making. We established and validated an easy, inexpensive, and rapid method for detecting DEL and L858R from cytologic material by high-resolution melting analysis (HRMA). Dilution for sensitivity studies revealed that DEL and L858R were detectable in the presence of at least 10% and 0.1% EGFR-mutant cells, respectively. We analyzed 37 archived cytological slides of specimens from 29 patients with advanced NSCLC and compared the results with direct sequencing data obtained previously. Of 37 samples, 34 (92%) yielded consistent results with direct sequencing, 2 were false negative, and 1 was indeterminate. The sensitivity of this analysis was 90% (19/21) and specificity, 100% (15/15). These results suggest that HRMA of archived cytologic specimens of advanced NSCLC is useful for detecting EGFR mutations in clinical practice.
authors.affiliation: Clinical Laboratory Division, National Cancer Center Hospital, Tokyo, Japan.
authors.forename: Kiyoaki
authors.initials: K
authors.lastname: Nomoto
authors.forename: Koji
authors.initials: K
authors.lastname: Tsuta
authors.forename: Toshimi
authors.initials: T
authors.lastname: Takano
authors.forename: Tomoya
authors.initials: T
authors.lastname: Fukui
authors.forename: Tomoya
authors.initials: T
authors.lastname: Fukui
authors.forename: Karin
authors.initials: K
authors.lastname: Yokozawa
authors.forename: Hiromi
authors.initials: H
authors.lastname: Sakamoto
authors.forename: Teruhiko
authors.initials: T
authors.lastname: Yoshida
authors.forename: Akiko Miyagi
authors.initials: AM
authors.lastname: Maeshima
authors.forename: Tatsuhiro
authors.initials: T
authors.lastname: Shibata
authors.forename: Koh
authors.initials: K
authors.lastname: Furuta
authors.forename: Yuichiro
authors.initials: Y
authors.lastname: Ohe
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Matsuno
mesh_terms: D001109:Archives; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D004252:DNA Mutational Analysis; D004273:DNA, Neoplasm / Q000235:genetics; D066246:ErbB Receptors / Q000235:genetics* / Q000378:metabolism; D017353:Gene Deletion; D018773:Genes, erbB-1; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
chemical_list: D004273:DNA, Neoplasm; D066246:ErbB Receptors