DNA methylation is a promising biomarker for forensic age estimation due to its strong correlation with chronological age. This study developed a droplet digital PCR (ddPCR) assay targeting three age-associated markers— ELOVL2, FHL2, and TRIM59—to evaluate its application in forensic cases. The assays were optimized using 100% methylated and unmethylated control DNA to evaluate probe specificity and sensitivity to methylation percentage. The assays exhibited high specificity, successfully distinguishing between methylated and unmethylated targets, and maintained sensitivity even at low methylation levels. Moreover, blood samples from three Korean individuals (aged 25, 42, and 55 years) were analyzed, revealing a clear age-related increase in methylation levels across all markers, with ELOVL2 exhibiting the strongest relation, followed by FHL2 and TRIM59. Reproducibility was evaluated by duplicate testing, with coefficient of variation values below 15% for all markers. These findings highlight the potential application of this ddPCR assay for forensic age estimation.
목차
Abstract Ⅰ. Introduction Ⅱ. Materials and Methods 1. DNA extraction and bisulfite conversion 2. Oligo design for quantification of methylation levels 3. Droplet Digital PCR analysis (ddPCR) Ⅲ. Results & Discussion 1. Specificity 2. Sensitivity to the methylation percentage 3. Assay assessment on 3 blood samples Ⅳ. Conclusion Ⅴ. Declaration of Competing Interest Ⅵ. Acknowledgment Ⅶ. References
키워드
Age predictionDNA methylationDroplet digital PCRForensic
저자
Rita Fakhr [ Department of Forensic Sciences, Sungkyunkwan University, Suwon, Republic of Korea ]
Un Na Koh [ Department of Forensic Sciences, Sungkyunkwan University, Suwon, Republic of Korea ]
So Eun Lee [ Department of Forensic Sciences, Sungkyunkwan University, Suwon, Republic of Korea ]
Si-Keun Lim [ Department of Forensic Sciences, Sungkyunkwan University, Suwon, Republic of Korea/ID-Cell Forensics Co., Sungkyunkwan University, Suwon, Republic of Korea/Convergence Bio Forensic Institute (CBFI), Biomedical Institute for Convergence at Sungkyunkwan University (BICS), Sungkyunkwan University, Suwon, Republic of Korea ]
Corresponding Author
법과학 분야는 사회정의 구현에 있어 크나큰 가치가 있음에도 불구하고 우리나라에서는 이 분야에 대한 인식이 미흡하여 선진 외국에 비해 침체되어 있는 실정이다. 이에 우리나라에서도 법과학 분야와 관련 있는 학계, 연구기관, 수사기관 등 유관 단체들로 구성된 한국 법과학회를 창립하여 이 분야를 활성화 시켜 과학수사를 한층 더 발전시키기 위함을 목적으로 한다.