Ultra-Deep Next-Generation Sequencing Of Plasma Cell-Free (Cf) Dna From Patients With Advanced Cancers

e22168 Background: Plasma cfDNA offers an easily obtainable source of DNA for molecular analysis, which provides an attractive alternative to tumor tissue testing. Novel ultrasensitive multiplex technologies are needed for further implementation of cfDNA testing in personalized cancer therapy. Methods: We have developed an ultra-deep next-generation sequencing method for detection of somatic alterations in 30 to 58 common cancer related genes in plasma cfDNA. Each cfDNA fragment was uniquely barcoded and amplified prior to Illumina target enrichment workflow, followed by ultra-deep sequencing ( u003e 10,000X). Proprietary data processing and analysis tools were developed to enable sensitive detection of rare mutant molecules over high wild-type background (detection of 1 in 1,000 - 10,000 molecules). Results were compared to molecular analysis of archival primary or metastatic tumor tissue obtained at different points of clinical care from a CLIA-certified laboratory. Results: Initially, cfDNA was extracted f...