Targeted Amplicon Sequencing Followed by Hybrid Selection Using Reference cfDNA: Variant Allele Frequency 0.1%
In this study, a novel target enrichment method called MAGNET-Seq was evaluated using genomic DNA samples with known variant allele frequencies (VAFs). The method combines two steps: (1) targeted multiplex PCR with unique molecular identifiers (UIDs), and (2) hybrid capture. Two primer sets were tested: a 43-target Drug Resistance Targeting Primers and a 7-target Reference Primers. Libraries were sequenced on an Illumina NovaSeq 6000 system. Performance was assessed by comparing observed vs known VAFs across 5 different levels (0%, 0.005%, 0.05%, 0.5%, 1%), and calculating sensitivity and specificity.