RRC ID 37765
著者 Uhara M, Matsuda K, Taira C, Higuchi Y, Okumura N, Yamauchi K.
タイトル Simple polymerase chain reaction for the detection of mutations and deletions in the epidermal growth factor receptor gene: applications of this method for the diagnosis of non-small-cell lung cancer.
ジャーナル Clin Chim Acta
Abstract BACKGROUND:Somatic mutations in the epidermal growth factor receptor (EGFR) gene are associated with the responses to the tyrosine kinase inhibitors gefitinib and erlotinib in patients with non-small-cell lung cancer (NSCLC). Although various methods for detecting EGFR gene mutations have been developed, they have several disadvantages. We attempted to establish a new method for the detection of EGFR gene mutations with the use of paraffin-embedded samples.
METHODS:The detections of T790M mutations in exon 20 and L858R mutations in exon 21 are based on the principle of allele-specific oligonucleotide polymerase chain reaction (PCR). We also designed PCR primers that enable to detect all types of deletions in exon 19. We assessed the basic performance efficiency of this method, and to confirm its clinical applicability, we performed PCR using DNA extracted from 66 tissue sections that were obtained from patients with NSCLC and embedded in paraffin.
RESULTS:The sensitivity of this method for the detection of deletions or mutations was as low as 0.5%. In the 66 subjects whose samples were analyzed, we detected the following deletions and mutations in the EGFR gene: 11 deletions in exon 19, 8 L858R mutations, and 1 double mutation of L858R and T790M.
CONCLUSION:The present method is sensitive and specific for the detection of deletions and mutations in the EGFR gene and is thus suitable for use in laboratory tests.
巻・号 401(1-2)
ページ 68-72
公開日 2009-3-1
DOI 10.1016/j.cca.2008.11.018
PII S0009-8981(08)00558-5
PMID 19063875
MeSH Carcinoma, Non-Small-Cell Lung / diagnosis* Carcinoma, Non-Small-Cell Lung / genetics Cell Line, Tumor DNA Mutational Analysis / methods* Exons / genetics Genes, erbB-1 / genetics* Humans Polymerase Chain Reaction / methods* Sensitivity and Specificity Sequence Deletion*
IF 2.615
引用数 12
WOS 分野 MEDICAL LABORATORY TECHNOLOGY
リソース情報
ヒト・動物細胞