Evaluation of a hybridization capture-based hereditary panel for forensic DNA analysis using next-generation sequencing.

Short tandem repeats (STR) remains the forensic gold standard genetic marker but face limitations with trace and degraded DNA. Single nucleotide polymorphism (SNP) offers significant potential to overcome these constraints. This study developed a novel hybridization capture-based SNP panel integrated with next-generation sequencing (NGS) for forensic DNA analysis. Using 8,464 unrelated Chinese Han individuals, we established population-specific allele frequencies for 2,998 biallelic SNPs (2,817 autosomal; 181 sex-chromosomal) after rigorous quality control. Exclusion of 19 SNPs deviating from Hardy-Weinberg equilibrium (HWE) and 8 SNPs in linkage disequilibrium (LD) yielded a final panel of 2,790 autosomal SNPs. The system demonstrated exceptional sensitivity, enabling reliable forensic analysis with 0.1 ng DNA input. Comparable genotyping performance was observed across diverse biological samples (blood cards, sperm stains, oral swabs, nails, hair roots) at 1-5 ng inputs. Although prolonged storage (3 months) reduced SNP recovery universally, buccal swabs exhibited more temporal stability than other substrates (p < 0.05). The panel achieved a total discrimination power (TDP) of 1-4.75 × 10 and cumulative probability of exclusion (CPE) of 1-4.61 × 10. Statistical simulations indicated that 31 SNPs suffice for identity determination, while 270 SNPs enable conclusive parentage testing in duo cases with 100 % sensitivity. This NGS-based solution provides robust forensic identification and kinship analysis, offering enhanced degradation tolerance, broad sample applicability, and population-tailored resolution for the Han Chinese cohort.