Methylation-Based Age Prediction Using Pyrosequencing Platform from Seminal Stains in Han Chinese Males

Various methods have been performed to predict an unknown individual's age from biological traces in forensic investigations. A considerably accurate age prediction for the semen donor can help narrow down the search in a sexual assault case. The aim of this study was to develop an assay for age prediction from seminal stains in Han Chinese males. We built a sperm-specific linear regression model using bisulfite pyrosequencing. Validations were conducted with a Mean Absolute Deviation from the chronological age (MAD) of 4.219 years in liquid semen, 4.158 years in fresh seminal stains, 4.393 years in aged seminal stains, and 3.880 years in mixed stains, respectively. Furthermore, our strategy enables accurate age prediction using a forensic casework sample. The strategy indicated that we produced an accurate and reliable age prediction tool for the semen donors in Han Chinese males for forensic purposes.