Abstract 4663: KRAS, BRAF, and EGFR mutational analysis in ovarian, colon, and lung cancers by highly multiplex PCR/barcoded-magnetic-bead (BMB) suspension-array assays

Abstract

Abstract Anti-epithelial growth factor receptor (EGFR) monoclonal antibodies and small molecule EGFR inhibitors have been used for treating metastatic colorectal cancer (mCRC) and non-small cell lung cancer (NSCLC), respectively. However, since a relatively large proportion of the mCRC and NSCLC patients carry the activating KRAS/BRAF mutations that can render these EGFR-targeted therapies ineffective, the KRAS/BRAF mutation testing is now required for such prescription. In contrast, epithelial ovarian cancer (EOC) thus far has been treated by surgery, radiotherapy, and/or chemotherapy depending on the tumor stages. However, it has been shown that a high percentage of tumors of the mucinous, endometrioid, low-grade serous, and other types of EOC patients also contain KRAS or BRAF somatic mutations, suggesting that drugs targeting the mediators of the EGFR pathway may have implications on treating EOC of certain types. The conventional analysis of the KRAS, BRAF, and EGFR mutations has been done by multiple reactions with one mutation target per reaction, thus requiring a large amount of precious patient sample for complete testing. Here we present three highly multiplex molecular diagnostic assays, utilizing amplification by allele-specific PCR and automatic detection by a platform designed around the barcoded-magnetic-bead (BMB) suspension-array technology, for detection of 12 KRAS (in codons 12 and 13), six BRAF (in codon 600), and 21 EGFR (in exons 18-21) mutations in mCRC, NSCLC, and EOC patient samples. The results indicate that, albeit all the reaction components are in single wells, these highly multiplex PCR/BMB assays, by comparing with the sequencing results, not only are sensitive and specific but also can conserve precious tissue specimens, save operating time and labor, reduce turnaround time, and increase assay throughput. Citation Format: Jason Lei, Julia Hsu, Peggy Jen, Daniel Huang, Andre Chung, Lloyd Kao, Miller Chang, Chiou-Chung Yuan, Wei-Hwa Lee, Chi-Tai Yeh, Dean Tsao. KRAS, BRAF, and EGFR mutational analysis in ovarian, colon, and lung cancers by highly multiplex PCR/barcoded-magnetic-bead (BMB) suspension-array assays. [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 4663. doi:10.1158/1538-7445.AM2014-4663