Detection of EGFR gene mutation in lung cancer by mutant-enriched polymerase chain reaction assay

Hiroaki Asano, Shinichi Toyooka, Masaki Tokumo, Kouichi Ichimura, Keisuke Aoe, Sachio Itou, Kazunori Tsukuda, Mamoru Oouchida, Motoi Aoe, Hideki Katayama, Akio Hiraki, Kazuro Sugi, Katsuyuki Kiura, Hiroshi Date, Nobuyoshi Shimizu

Research output: Contribution to journalArticle

167 Citations (Scopus)

Abstract

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 × 103 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.

Original languageEnglish
Pages (from-to)43-48
Number of pages6
JournalClinical Cancer Research
Volume12
Issue number1
DOIs
Publication statusPublished - Jan 1 2006

Fingerprint

erbB-1 Genes
Lung Neoplasms
Polymerase Chain Reaction
Mutation
Epidermal Growth Factor Receptor
Tomography
Non-Small Cell Lung Carcinoma
Lung
Neoplasms
Genes
Biopsy
Needle Biopsy
Protein-Tyrosine Kinases
Digestion
Exons
Research Design
Biomarkers
Enzymes

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Detection of EGFR gene mutation in lung cancer by mutant-enriched polymerase chain reaction assay. / Asano, Hiroaki; Toyooka, Shinichi; Tokumo, Masaki; Ichimura, Kouichi; Aoe, Keisuke; Itou, Sachio; Tsukuda, Kazunori; Oouchida, Mamoru; Aoe, Motoi; Katayama, Hideki; Hiraki, Akio; Sugi, Kazuro; Kiura, Katsuyuki; Date, Hiroshi; Shimizu, Nobuyoshi.

In: Clinical Cancer Research, Vol. 12, No. 1, 01.01.2006, p. 43-48.

Research output: Contribution to journalArticle

Asano, Hiroaki ; Toyooka, Shinichi ; Tokumo, Masaki ; Ichimura, Kouichi ; Aoe, Keisuke ; Itou, Sachio ; Tsukuda, Kazunori ; Oouchida, Mamoru ; Aoe, Motoi ; Katayama, Hideki ; Hiraki, Akio ; Sugi, Kazuro ; Kiura, Katsuyuki ; Date, Hiroshi ; Shimizu, Nobuyoshi. / Detection of EGFR gene mutation in lung cancer by mutant-enriched polymerase chain reaction assay. In: Clinical Cancer Research. 2006 ; Vol. 12, No. 1. pp. 43-48.
@article{37cdfc5a7b614552bb0782cb4d3bf0fd,
title = "Detection of EGFR gene mutation in lung cancer by mutant-enriched polymerase chain reaction assay",
abstract = "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 × 103 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.",
author = "Hiroaki Asano and Shinichi Toyooka and Masaki Tokumo and Kouichi Ichimura and Keisuke Aoe and Sachio Itou and Kazunori Tsukuda and Mamoru Oouchida and Motoi Aoe and Hideki Katayama and Akio Hiraki and Kazuro Sugi and Katsuyuki Kiura and Hiroshi Date and Nobuyoshi Shimizu",
year = "2006",
month = "1",
day = "1",
doi = "10.1158/1078-0432.CCR-05-0934",
language = "English",
volume = "12",
pages = "43--48",
journal = "Clinical Cancer Research",
issn = "1078-0432",
publisher = "American Association for Cancer Research Inc.",
number = "1",

}

TY - JOUR

T1 - Detection of EGFR gene mutation in lung cancer by mutant-enriched polymerase chain reaction assay

AU - Asano, Hiroaki

AU - Toyooka, Shinichi

AU - Tokumo, Masaki

AU - Ichimura, Kouichi

AU - Aoe, Keisuke

AU - Itou, Sachio

AU - Tsukuda, Kazunori

AU - Oouchida, Mamoru

AU - Aoe, Motoi

AU - Katayama, Hideki

AU - Hiraki, Akio

AU - Sugi, Kazuro

AU - Kiura, Katsuyuki

AU - Date, Hiroshi

AU - Shimizu, Nobuyoshi

PY - 2006/1/1

Y1 - 2006/1/1

N2 - 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 × 103 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.

AB - 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 × 103 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.

UR - http://www.scopus.com/inward/record.url?scp=31544459265&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=31544459265&partnerID=8YFLogxK

U2 - 10.1158/1078-0432.CCR-05-0934

DO - 10.1158/1078-0432.CCR-05-0934

M3 - Article

C2 - 16397022

AN - SCOPUS:31544459265

VL - 12

SP - 43

EP - 48

JO - Clinical Cancer Research

JF - Clinical Cancer Research

SN - 1078-0432

IS - 1

ER -