3 Results
3.1 Selection of sequencing methods for neonatal screening
To select the appropriate detection method, we firstly compared different sequencing methods for NBS used in the previously published studies (Table 1). In the beginning, scientists conducted neonatal screening trials by using whole exome sequencing (WES) or whole genome sequencing (WGS), including WGS for STATseq project16, WES for BabySeq project17, NBSeq project27, and North Carolina Newborn Exome Sequencing for Universal Screening (NC NEXUS) project18. These studies implied sequencing may screen out inborn errors before the disease onset and WGS/WES is operationally feasible in neonatal screening28-32. However, WGS/WES analysis produces lots of VUS, shows the higher cost and longer turn-around time (TAT). Consequently, scientists explored the application of targeted NGS panels that only analyzed a subset of gene loci in NBS. Notably, studies suggested that targeted sequencing has more advantages over WGS/WES as the operation of data analysis/follow-up is more feasible and easier, with lower cost, shorter TAT, and easier interpretation33,34. Additionally, there is little difference in positive rate between targeted NGS and WGS/WES35. Therefore, we choose targeted sequencing in our study.