A mutation-sensitive switch assay to detect five clinically significant epidermal growth factor receptor mutations.
Epidermal growth factor receptor (EGFR) mutations can affect the therapeutic efficacy of drugs used to treat nonsmall-cell lung cancer (NSCLC). We aimed to develop methods to detect five common EGFR somatic mutations in tumor tissues from NSCLC patients by using a nanoscale mutation-sensitive switch consisting of a high-fidelity polymerase and phosphorothioate-modified allele-specific primers. The five clinically significant EGFR mutations examined here are S768I, T790M, L858R, and 15- and 18-bp deletion mutations in exon 19. Our assays showed sensitivities of 100 copies and specificities of more than three log scales for matched templates relative to mismatched templates by routine polymerase chain reaction (PCR), real-time PCR, and multiplex PCR. This assay would be superior to DNA sequencing in situations where mutant DNA is not abundant.
Department of Molecular Diagnostics and Biopharmaceutics, College of Pharmaceutical Science, Soochow University , Suzhou, China .
Bin
B
Liu
Department of Molecular Diagnostics and Biopharmaceutics, College of Pharmaceutical Science, Soochow University , Suzhou, China .
Lin
L
Zhou
Department of Molecular Diagnostics and Biopharmaceutics, College of Pharmaceutical Science, Soochow University , Suzhou, China .
Qian
Q
Wang
Department of Molecular Diagnostics and Biopharmaceutics, College of Pharmaceutical Science, Soochow University , Suzhou, China .
Kai
K
Li
D001681:Biological Assay; D002289:Carcinoma, Non-Small-Cell Lung / Q000097:blood / Q000235:genetics* / Q000628:therapy; D016022:Case-Control Studies; D017931:DNA Primers; D066246:ErbB Receptors / Q000097:blood / Q000235:genetics*; D005091:Exons; D005812:Genes, Switch; D018773:Genes, erbB-1; D006580:Genetic Carrier Screening / Q000379:methods*; D006801:Humans; D008175:Lung Neoplasms / Q000097:blood / Q000235:genetics* / Q000628:therapy; D060888:Real-Time Polymerase Chain Reaction; D017384:Sequence Deletion
D017931:DNA Primers; C512478:EGFR protein, human; D066246:ErbB Receptors