Comparison Test of Sensitivity Between Next Generation Sequencing (NGS) Hotspot Panel and Droplet Digital PCR of KRAS G12 / G13 Mutation

Abstract

Cancer is an abnormal proliferation of cells that is characterized by the presence of genomic alterations including DNA mutations, deletions, insertions, translocations, inversions, and others. KRAS gene is one of the most mutated genes across different cancer types. The most common mutations in KRAS are found in codons 12 and 13 of KRAS protein, which are associated with a lack of response to anti- epidermal growth factor receptor (EGFR) antibody therapy. This study assessed and compared the performance between two diagnostic methods: droplet digital PCR (ddPCR) and next generation sequencing (NGS). The main goal was to determine KRAS G12 / G13 mutant allele fraction using NGS and to compare the accuracy toddPCR. A total of 28 samples of non – small cell lung cancer (NSCLC) and colorectal cancer (CRC) were analyzed using ddPCR and NGS methods. Our results show that even though both methods exhibited high rate of concordance and correlation, the study proved that ddPCR is more superior when it comes to detecting low frequency mutations. Even though strong correlation was observed, based on the values obtained, we concluded that ddPCR is more accurate, reliable, and sensitive in comparison with NGS.