Y-CpG-based semen age prediction: analysis of vasectomized samples and development of an optimized multiplex assay evaluated in independent and mixed samples.

Λεπτομέρειες βιβλιογραφικής εγγραφής
Τίτλος:	Y-CpG-based semen age prediction: analysis of vasectomized samples and development of an optimized multiplex assay evaluated in independent and mixed samples.
Συγγραφείς:	Lee JE; Department of Forensic Medicine, Seoul National University College of Medicine, 103 Daehak-Ro, Jongno-Gu, Seoul, 03080, South Korea., Lee HY; Department of Forensic Medicine, Seoul National University College of Medicine, 103 Daehak-Ro, Jongno-Gu, Seoul, 03080, South Korea. hylee192@snu.ac.kr.; Institute of Forensic and Anthropological Science, Seoul National University College of Medicine, 103 Daehak-Ro, Jongno-Gu, Seoul, 03080, South Korea. hylee192@snu.ac.kr.
Πηγή:	International journal of legal medicine [Int J Legal Med] 2026 Jul; Vol. 140 (4), pp. 2055-2064. Date of Electronic Publication: 2026 Apr 14.
Τύπος έκδοσης:	Journal Article
Γλώσσα:	English
Στοιχεία περιοδικού:	Publisher: Springer International Country of Publication: Germany NLM ID: 9101456 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1437-1596 (Electronic) Linking ISSN: 09379827 NLM ISO Abbreviation: Int J Legal Med Subsets: MEDLINE
Imprint Name(s):	Original Publication: Heidelberg, FRG : Springer International, c1990-
Ιατρικοί όροι (MeSH):	Semen/chemistry , Aging/genetics , Chromosomes, Human, Y* , DNA Methylation* , CpG Islands* , Vasectomy*, Forensic Genetics/methods ; Cervix Mucus/chemistry ; Humans ; Male ; Adult ; Female ; Multiplex Polymerase Chain Reaction ; Middle Aged
Περίληψη:	Age prediction using DNA methylation has become an important tool in forensic genetics, especially for semen samples in sexual assault cases. Our previous research has identified age-associated CpG markers on the Y-chromosome and developed a multiplex assay along with regression models for predicting the age of semen samples. However, the performance of these methods under biologically altered conditions, such as vasectomized semen and mixed body fluid samples, has not been thoroughly investigated. In this study, we analyzed semen samples from vasectomized individuals using a previously developed 10-amplicon Y-CpG assay to examine methylation patterns and age prediction capabilities. The vasectomized samples were categorized based on body fluid identification results, revealing distinct methylation profiles. We observed markedly reduced age prediction accuracy in samples that lacked semen-specific profiles, while those with semen-specific profiles exhibited patterns similar to normal semen. Building on feature selection outcomes from our earlier study, we further developed an optimized five-amplicon multiplex assay and assessed its performance with independent semen samples and semen-vaginal fluid mixtures. In evaluations involving 11 independent semen samples, the stepwise and lasso models produced mean absolute errors (MAEs) of 6.74 and 6.55 years, respectively. The assay also showed consistent performance with semen-vaginal fluid mixtures, with methylation differences below 2% and average age prediction errors under 6 years. Overall, these findings indicate that the five-amplicon assay offers a streamlined and practical approach for predicting semen age in forensic contexts. Additionally, incorporating body fluid identification may enhance the interpretation of age predictions in challenging samples.
Competing Interests:	Declarations. Ethics approval: This study was approved by the Institutional Review Board of Yonsei University Severance Hospital (IRB No. 4–2015-0083) and Seoul National University Hospital (IRB No. 1912–053-1087), in accordance with the Declaration of Helsinki and ICH-GCP (International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use – Good Clinical Practice). Consent to participate: Written informed consent was obtained from all participants prior to sample collection. Clinical trial number: Not applicable. Consent for publication: Not applicable. Competing interest: The authors have declared no conflict of interest.
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Contributed Indexing:	Keywords: Age; Massively parallel sequencing; Methylation; Semen; Y-chromosome
Entry Date(s):	Date Created: 20260414 Date Completed: 20260628 Latest Revision: 20260726
Update Code:	20260726
PubMed Central ID:	PMC13275618
DOI:	10.1007/s00414-026-03797-y
PMID:	41979672
Βάση Δεδομένων:	MEDLINE

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ISSN:	1437-1596
DOI:	10.1007/s00414-026-03797-y