The Use Of Improved And Complete Enrichment Co-Amplification At Lower Denaturation Temperature (Ice Cold-Pcr) Method For The Detection Of Egfr And Kras Mutations From Cell-Free Plasma Dna Of Non-Small Cell Lung Cancer (Nsclc) Patients

8058 Background: Identification of specific molecular alterations from cell free plasma DNA (cfpDNA) holds tremendous potential as a noninvasive method to assess tumor genotype. We evaluated whether ICE COLD-PCR(ICP) can be used to identify EGFR and KRAS mutations from cfpDNA in patients enrolled in the BATTLE research clinical trial. Methods: Tissue genotyping of KRAS (Exons 2, 3) and EGFR (Exons 18 - 21) on DNA extracted form paraffin-embedded tumor tissue was determined using PCR-based sequencing analysis, with lower limit of sensitivity of detection of 20%. Genotyping of cfpDNA was determined using ICP for mutation enrichment followed by Sanger sequencing for mutation detection, with limit of detection of 0.05%. Results: DNA was isolated and extracted from 154 available plasma samples with matched tumor genotype; with volumes ranging 0.2–0.7 mL. For the overall population with or without mutations, a concordance of 92%, 91%, and 81% was observed with tissue genotypes for EGFR Exon 19, EGFR Exon 21 and...