Sriwijaya Journal of Forensic and Medicolegal
Vol. 3 No. 1 (2025): Sriwijaya Journal of Forensic and Medicolegal

Reading the Epigenetic Clock: A Comparative Analysis of DNA Methylation Markers for Age Estimation in Semen, Saliva, and Touch DNA

Febria Suryani (Unknown)
Bryan Helsey (Unknown)
Leonardo Simanjuntak (Unknown)
Karina Chandra (Unknown)
Mustafa Mahmud (Unknown)
Lisha Sandrina (Unknown)
Ahmad Erza (Unknown)



Article Info

Publish Date
24 Jul 2025

Abstract

Introduction: The capacity to predict an individual's age from biological evidence constitutes a significant advancement in forensic intelligence. DNA methylation, a stable epigenetic mark, provides a molecular basis for "epigenetic clocks." However, the operational reliability of these clocks necessitates rigorous validation across diverse biological samples and populations, particularly for challenging, low-template touch DNA evidence. Methods: Following approval from the Ethical Committee of CMHC Indonesia (No. 128/EC/CMHC/2023), we recruited 150 healthy Indonesian male volunteers aged 18-65. Semen, saliva, and high-yield standardized touch DNA samples were collected. DNA was extracted, quantified fluorometrically, and subjected to bisulfite conversion with efficiency controls. The methylation levels of a curated five-CpG panel (ELOVL2, FHL2, TRIM59, KCNQ1DN, C1orf132) were quantified using a rigorously controlled pyrosequencing workflow. Body-fluid-specific age prediction models were developed using multiple linear regression, validated with 10-fold cross-validation, and assessed for statistical assumptions including multicollinearity. Results: The models for semen and saliva demonstrated high predictive accuracy, yielding Mean Absolute Deviation (MAD) values of 3.19 years (R²=0.94) and 3.55 years (R²=0.92), respectively. The model developed from high-yield touch DNA was less precise but still highly informative, with a MAD of 5.49 years (R²=0.85). All models satisfied the assumptions of linear regression, with Variance Inflation Factors below 2.5 indicating low multicollinearity. The 95% prediction intervals were narrowest for semen, reflecting its superior precision. Conclusion: This study validates a robust, targeted epigenetic panel for age prediction in a Southeast Asian population. We present highly accurate, tissue-specific models for semen and saliva, suitable for immediate consideration in forensic casework. The touch DNA model, while requiring cautious interpretation, provides a valuable framework for generating investigative leads from trace evidence. Our findings underscore the critical importance of tissue-specific modeling and provide a detailed methodological and statistical blueprint for the responsible implementation of forensic age estimation.

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Journal Info

Abbrev

SJFM

Publisher

Subject

Biochemistry, Genetics & Molecular Biology Health Professions Immunology & microbiology Law, Crime, Criminology & Criminal Justice Medicine & Pharmacology

Description

Focus Sriwijaya Journal of Forensic and Medicolegal (SJFM) focused on the development of medical sciences especially forensic and medicolegal for human well-being. Scope Sriwijaya Journal of Forensic and Medicolegal (SJFM) publishes articles which encompass all aspects of basic research/clinical ...