使用SureSelect Cancer Pan Heme检测对血液系统恶性肿瘤进行整合式长读段靶标富集和全面基因组分析

Next-generation sequencing (NGS) has transformed cancer research, yet its effectiveness is often constrained by sample quality, tumor fraction, and limitations inherent to short-read sequencing-particularly in detecting structural variants, complex rearrangements, and alterations in repetitive or polymorphic regions. Long-read sequencing offers a solution, but cost and throughput challenges persist. To address these limitations, we developed a flexible, automation-compatible library preparation and target enrichment workflow that supports DNA inputs down to 200 ng, accommodates both enzymatic and mechanical shearing, and enables fast hybridization (90 minutes). Coupled with long-read sequencing and the SureSelect Cancer Pan Heme assay, this platform enables detection of main genomic alterations-including SNVs, indels, CNVs, and gene fusions-within a single assay. The SureSelect Cancer Pan Heme panel, codeveloped with Roswell Park Comprehensive Cancer Center, interrogates 359 DNA and 124 RNA genes, delivering integrated DNA/RNA analysis and overcoming the limitations of conventional single-analyte methods such as karyotyping, FISH, and PCR. Although in this study, we only focus on DNA as the substrate. We demonstrate high enrichment efficiency using a novel fast hybridization buffer, achieving on-target rates >80% for libraries with insert sizes up to 4-5 kb. Comparative analysis versus short-read approaches reveals superior coverage in challenging genomic regions using enriched long-read sequencing as well as allowing to phase variants. This solution offers a cost-effective, scalable, and rapid-turnaround workflow for molecular laboratories, advancing precision oncology in hematologic malignancies. For Research Use Only. Not for use in diagnostic procedures.