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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2006-6-16
pubmed:abstractText
The detection of mutations in the epidermal growth factor receptor (EGFR) gene impacts therapeutic decision-making for non-small-cell lung carcinoma (NSCLC). Although direct sequencing has been most frequently used to detect EGFR mutations, this method displays several disadvantages. We set up simple mutation-specific polymerase chain reaction (PCR) for common delE746-A750 and L858R mutations of EGFR using primers specific to these mutations. Both mutation-specific PCR and direct sequencing methods were used to investigate 62 samples of NSCLC, and the results were compared. To evaluate the sensitivity of mutation-specific PCR, DNA mixtures containing various proportions of mutant alleles were analyzed. Mutation-specific PCR revealed delE746-A750 in 8 samples and L858R in 14 samples. All samples with either delE746-A750 or L858R mutation revealed by direct sequencing also displayed positive results for mutation-specific PCR. Conversely, mutations in 3 samples revealed by L858R-specific PCR were barely detectable by direct sequencing. In DNA mixture analysis, DNA mixtures containing 2.5% of delE746-A750 allele or 0.25% of L858R allele yielded positive results with mutation-specific PCR. Our mutation-specific PCR showed satisfactory sensitivity and reliability for detecting major EGFR mutations in clinical NSCLC samples. Given the practical availability, this method could be widely applicable to the treatment of lung cancers.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jun
pubmed:issn
1052-9551
pubmed:author
pubmed:issnType
Print
pubmed:volume
15
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
101-8
pubmed:dateRevised
2009-11-19
pubmed:meshHeading
pubmed-meshheading:16778591-Base Sequence, pubmed-meshheading:16778591-Carcinoma, Non-Small-Cell Lung, pubmed-meshheading:16778591-Case-Control Studies, pubmed-meshheading:16778591-Cell Line, Tumor, pubmed-meshheading:16778591-DNA, Neoplasm, pubmed-meshheading:16778591-DNA Mutational Analysis, pubmed-meshheading:16778591-DNA Primers, pubmed-meshheading:16778591-Humans, pubmed-meshheading:16778591-Lung Neoplasms, pubmed-meshheading:16778591-Mutation, pubmed-meshheading:16778591-Point Mutation, pubmed-meshheading:16778591-Polymerase Chain Reaction, pubmed-meshheading:16778591-Protein Structure, Tertiary, pubmed-meshheading:16778591-Protein-Tyrosine Kinases, pubmed-meshheading:16778591-Receptor, Epidermal Growth Factor, pubmed-meshheading:16778591-Sensitivity and Specificity, pubmed-meshheading:16778591-Sequence Deletion
pubmed:year
2006
pubmed:articleTitle
A simple and sensitive method for detecting major mutations within the tyrosine kinase domain of the epidermal growth factor receptor gene in non-small-cell lung carcinoma.
pubmed:affiliation
Department of Laboratory Medicine, Kyorin University, Tokyo, Japan. onishi@kyorin-u.ac.jp
pubmed:publicationType
Journal Article, Comparative Study