The use of COLD-PCR and pyrosequencing for sensitive detection of EGFR T790M mutation

A sensitive and convenient method for the detection of epidermal growth factor receptor (EGFR) T790M mutation in non-small cell lung cancer (NSCLC) patients with acquired resistance to tyrosine kinase inhibitors (TKIs) would be desirable guide treatment strategy. Consequently, studies have focused on characterization EGFR mutation. Herein, two methods co-amplification at lower denaturation temperature PCR (COLD-PCR) pyrosequencing were combined (COLDPCR/ pyrosequencing) detecting Evaluation mutation-containing dilutions revealed that sensitivities COLD-PCR/pyrosequencing conventional PCR/pyrosequencing assays 0.1 5%, respectively, indicating a 50-fold increase sensitivity. When 20 clinical NSCLC samples who had relapsed under firstgeneration TKI further determined using PCR/pyrosequencing, rates 35% (7/20) 25% (5/20), respectively. All positive also found COLD-PCR/pyrosequencing. The discordant cases 2 no detected but which is cost-effective tool will permit an improvement therapeutic management.