Detection of EGFR Gene Mutation in Lung Cancer by Mutant-Enriched Polymerase Chain Reaction Assay

• Published in: Clinical cancer research Vol. 12; no. 1; pp. 43 - 48
• Main Authors: ASANO, Hiroaki, TOYOOKA, Shinichi, HIRAKI, Akio, SUGI, Kazuro, KIURA, Katsuyuki, DATE, Hiroshi, SHIMIZU, Nobuyoshi, TOKUMO, Masaki, ICHIMURA, Kouichi, AOE, Keisuke, ITO, Sachio, TSUKUDA, Kazunori, OUCHIDA, Mamoru, AOE, Motoi, KATAYAMA, Hideki
• Format: Journal Article
• Language: English
• Published: Philadelphia, PA American Association for Cancer Research 01.01.2006
• Subjects: Antineoplastic agents; Biological and medical sciences; Biopsy; Carcinoma, Non-Small-Cell Lung - genetics; Carcinoma, Non-Small-Cell Lung - pathology; CT-guided needle lung biopsy; DNA Mutational Analysis; DNA Primers; EGFR; Female; Genes, erbB-1 - genetics; Humans; Lung Neoplasms - genetics; Lung Neoplasms - pathology; Medical sciences; mutant-enriched PCR; Mutation; NSCLC; Pharmacology. Drug treatments; Pleural Effusion; pleural fluid; Pneumology; Polymerase Chain Reaction - methods; Sensitivity and Specificity; Tumors of the respiratory system and mediastinum; Lung disease; Respiratory disease; Lung cancer; Malignant tumor; Bronchopulmonary; Epidermal growth factor receptor; Polymerase chain reaction; Gene; Bronchus disease; Genetics; Diagnosis; Mutation; Molecular biology; Detection
• DOI: 10.1158/1078-0432.CCR-05-0934
• ISSN: 1078-0432

Purpose: Mutations in the epidermal growth factor receptor ( EGFR ) gene have been reported to be present in non–small cell lung cancer (NSCLC) and related to the responsiveness of tumors to EGFR tyrosine kinase inhibitors, suggesting its usefulness as a biomarker. Because clinical samples contain tumor and normal cells or genes, a highly sensitive assay for detecting mutation is critical for clinical applications. Experimental Design: The mutant-enriched PCR is a rapid and sensitive assay with selective restriction enzyme digestion. We developed the mutant-enriched PCR assay targeting exons 19 and 21 of EGFR and applied the developed assay to detect mutations in 108 cases of surgically resected specimens of NSCLCs, 18 samples of computed tomography (CT)–guided needle lung biopsies, and 20 samples of pleural fluid. In addition, results were then compared with those from direct sequencing and a nonenriched PCR assay. Results: The mutant-enriched PCR that was proved to enrich one mutant of 2 × 10 3 normal genes detected mutations in 37 cases of 108 resected tumors, seven samples of CT-guided lung biopsies, and seven samples of pleural fluid. Among mutant cases, four resected tumors, two CT-guided lung biopsies, and two pleural fluid were identified as additional mutant cases by the mutant-enriched PCR, which were considered normal based on nonenriched assays. Conclusions: Our results indicate that EGFR mutations are readily detectable by mutant-enriched PCR in various clinical samples. Thus, mutant-enriched PCR may provide a valuable method of potentially detecting a small fraction of mutant genes in heterogeneous specimens, indicating its possible use in clinical application for NSCLC.