Abstract 1509: A ARMS-PNA PCR assay for detection of EGFR mutations in FFPE tumor samples

Abstract

Abstract The activating mutations of Epidermal growth factor receptor (EGFR) are the predictive markers for response to tyrosine kinase inhibitors gefitinib and erlotinib in non-small cell lung cancer(NSCLC). Therefore detection of EGFR mutations will guide effective therapeutical options for patients with NSCLC. Here we developed a new method for the rapid and reliable detection of EGFR mutations by using a combination of modified mutation-specific ARMS primers and peptide nucleic acid (PNA) clamping system, named ARMS-PNA assay. In this real-time PCR system, the wild-type EGFR DNA is completely blocked by PNA with a higher annealing temperature, and the mutant gene is selectively amplified by the modified ARMS primers and the amplicon extending signals are collected by the taqman-probe at the same time. The ARMS-PNA assay could detect 29 mutations on EGFR gene with a sensitivity of 0.5% under the background of a total of 10ng wild-type genomic DNA. By using this assay, we analyzed the EGFR mutations in 1033 Chinese NSCLC FFPE samples in comparison with Sanger sequencing. Our results showed that the positive rates were 50.1%, which were higher than that measured by Sanger sequencing (48.3%). The coincidence rates of the two methods were 99.0% for the positive samples and 95.5% for the negative samples. The total coincidence rate was 97.2%. In conclusion, our study provides a basis for the detection of somatic mutations in EGFR in a routine clinical setting. The ARMS-PNA PCR assay allows the detection of the activating EGFR mutations in FFPE samples with better sensitivity and feasibility than the current assays used in clinic.<!–EndFragment–> Citation Format: Shanshan Zhao, Xiao Guo, Ziwen Long, Dehua Yu. A ARMS-PNA PCR assay for detection of EGFR mutations in FFPE tumor samples. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1509. doi:10.1158/1538-7445.AM2014-1509