MSIplus for Integrated Colorectal Cancer Molecular Testing by Next-Generation Sequencing.

摘要: Molecular analysis of colon cancers currently requires multiphasic testing that uses various assays with different performance characteristics, adding cost and time to patient care. We have developed a single, next-generation sequencing assay simultaneously evaluate colorectal for mutations in relevant cancer genes (KRAS, NRAS, BRAF) tumor microsatellite instability (MSI). In sample set 61 cases, the demonstrated overall sensitivity 100% specificity identifying cancer-associated mutations, practical limit detection at 2% mutant allele fraction. MSIplus was 97% sensitive (34 35 MSI-positive cases) specific (42 42 MSI-negative ascertaining MSI phenotype cohort 78 specimens. These characteristics were slightly better than conventional multiplex PCR (97% 95% specificity), which is based on comparison loci amplified from matched normal material, applied same specimen cohort. Because an amplicon approach, it rapid appropriate specimens limited available material or fragmented DNA. This integrated strategy offers several advantages over existing methods, including lack need unbiased variants target genes, automated pipeline enabling principled reproducible identification status simultaneously.