RHD-RHCE high fidelity long-read sequencing

We sequenced the genomic region encompassing the RHD and RHCE human blood group genes. The Rh blood group is the second most clinically significant blood group system in human transfusion, following the ABO system. However, due to the high sequence similarity between RHD and RHCE, reliably sequencing either gene remains highly challenging. As a result, 1) full-length reference sequences for these two genes are rarely available, and 2) previous NGS approaches often introduced significant errors during the mapping and variant calling processes.Long-read sequencing has facilitated the reliable assembly of this highly complex target region by leveraging its extended read lengths. Following target enrichment with 32 extensively overlapping amplicons, each spanning 25 kb, we sequenced a total of 170 kb of the target region using PacBio HiFi sequencing. This approach enabled us to completely phase-resolve the entire RHD-RHCE region and generate a comprehensive catalog of RHD-RHCE reference sequences across 63 individuals.