Rapid polymerase chain reaction-based detection of epidermal growth factor receptor gene mutations in lung adenocarcinoma Journal Article
| Authors: | Pan, Q.; Pao, W.; Ladanyi, M. |
| Article Title: | Rapid polymerase chain reaction-based detection of epidermal growth factor receptor gene mutations in lung adenocarcinoma |
| Abstract: | Somatic mutations in the tyrosine kinase domain of the epidermal growth factor receptor (EGFR) gene are present in lung adenocarcinomas that respond to the EGFR inhibitors gefitinib and erlotinib. Two types of mutations account for ∼90% of mutated cases: short in-frame deletions in exon 19 and a specific point mutation in exon 21 at codon 858 (L858R). Screening for these mutations has been based mainly on direct sequencing. We report here the development and validation of polymerase chain reaction-based assays for these two predominant types of EGFR mutations. The assay for exon 19 mutations is based on length analysis of fluorescently labeled polymerase chain reaction products, and the assay for the exon 21 L858R mutation is based on a new Sau96I restriction site created by this mutation. Using serial dilutions of DNAs from lung cancer cell lines harboring either exon 19 or 21 mutations, we detected these mutations in the presence of up to ∼90% normal DNA. In a test set of 39 lung cancer samples, direct sequencing detected mutations in 25 cases whereas our assays were positive in 29 cases, including 4 cases In which mutations were not apparent by sequencing. These assays offer higher sensitivity and ease of scoring and eliminate the need for sequencing, providing a robust and accessible approach to the rapid identification of most lung cancer patients likely to respond to EGFR inhibitors. Copyright © American Society for Investigative Pathology and the Association for Molecular Pathology. |
| Keywords: | clinical article; controlled study; human tissue; somatic mutation; exon; mutation; exons; sequence deletion; validation process; polymerase chain reaction; sensitivity analysis; adenocarcinoma; lung neoplasms; genotype; receptor, epidermal growth factor; cancer cell culture; mutational analysis; lung adenocarcinoma; dna; amino acid sequence; molecular sequence data; drug response; dna, neoplasm; scoring system; intermethod comparison; base sequence; dna sequence; dna mutational analysis; codon; point mutation; genetic screening; fluorescence analysis; epidermal growth factor receptor kinase; deletion mutant; epidermal growth factor receptor kinase inhibitor; restriction site |
| Journal Title: | Journal of Molecular Diagnostics |
| Volume: | 7 |
| Issue: | 3 |
| ISSN: | 1525-1578 |
| Publisher: | Elsevier Science, Inc. |
| Date Published: | 2005-08-01 |
| Start Page: | 396 |
| End Page: | 403 |
| Language: | English |
| PUBMED: | 16049312 |
| PROVIDER: | scopus |
| PMCID: | PMC1867537 |
| DOI: | 10.1016/s1525-1578(10)60569-7 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 116" - "Export Date: 24 October 2012" - "CODEN: JMDIF" - "Source: Scopus" |
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