Developing a DNA methylation assay for human age prediction in blood and bloodstain |
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| Institution: | 1. Institute of Forensic Medicine, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, Sichuan, China;2. Clinical and Translational Research Institute, University of California, San Diego. 9500 Gilman Dr., MC 0990, La Jolla, CA 92093-0990, USA;1. Department of Chemistry and Biochemistry, Florida International University, Miami, FL, USA;2. General Department of Forensic Science and Criminology, Dubai Police, Dubai, United Arab Emirates;3. Faculty of Biosciences, Laboratory of Human and Molecular Genetics, PUCRS, Porto Alegre, Brazil,;4. School of Criminal Justice, University of Southern Mississippi, Hattiesburg, MS, USA;1. Department of Genetic Identification, Erasmus MC University Medical Center Rotterdam, Rotterdam, The Netherlands;2. Key Laboratory of Genomic and Precision Medicine, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, China;3. Department of Internal Medicine, Erasmus MC University Medical Center Rotterdam, Rotterdam, The Netherlands;4. Center for Biomics, Erasmus MC University Medical Center Rotterdam, Rotterdam, The Netherlands;5. Department of Epidemiology, Erasmus MC University Medical Center Rotterdam, Rotterdam, The Netherlands;6. Epigenomics AG, Berlin, Germany;1. King’s Forensics, Department of Analytical, Environmental and Forensic Sciences, Faculty of Life Sciences and Medicine, King’s College London, 150 Stamford Street, London SE1 9NH, United Kingdom;2. Centre for Forensic Science, Department of Applied Sciences, Faculty of Health and Life Sciences, Northumbria University Newcastle, Ellison Building, NE1 8ST Newcastle Upon Tyne, United Kingdom;1. University of Amsterdam, Swammerdam Institute for Life Sciences, Science Park 904, 1098XH Amsterdam, The Netherlands;2. Netherlands Forensic Institute, Biological Traces, Laan van Ypenburg 6, 2497GB Den Haag, The Netherlands;3. University of Amsterdam, Institute for Biodiversity and Dynamics, Science Park 904, 1098XH Amsterdam, The Netherlands;1. Forensic Genetics Unit, Institute of Forensic Sciences, University of Santiago de Compostela, Spain;2. Faculty of Mathematics, University of Santiago de Compostela, Spain;3. Spanish National Genotyping Center-USC-PRB2-ISCIII, Santiago de Compostela, Spain;4. Institute of Legal Medicine, Faculty of Medicine, University of Cologne, Cologne, Germany;5. Institute of Zoology, Faculty of Biology and Earth Sciences, Jagiellonian University, Krakow, Poland;6. Malopolska Centre of Biotechnology, Jagiellonian University, Krakow, Poland;7. Center of Excellence in Genomic Medicine Research, King Abdulaziz University, Jeddah, Saudi Arabia;1. Epigenomics Research Center, Genome Institute, Korea Research Institute of Bioscience and Biotechnology, 125 Gwahak-ro, Yuseong-gu, Daejeon, Republic of Korea;2. Department of Functional Genomics, University of Science of Technology, 125 Gwahak-ro, Yuseong-gu, Daejeon, Republic of Korea;3. DNA Forensic Division, Supreme Prosecutor’s Office, 157 Banpodae-ro, Seocho-gu, Seoul, Republic of Korea |
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| Abstract: | Age prediction of an individual based on biological traces remained in a crime scene is of ultimate importance for criminal investigation. Growing evidence indicates that some CpG sites may have age-related methylation changes and thus may be a promising tool for age prediction. In this study, we utilized the pyrosequencing approach to screen age-related CpG (AR-CpG) sites for age prediction. Five AR-CpGs were identified as age-related markers from thirty-eight candidates, among which three CpG sites, ITGA2B_1, NPTX2_3, and NPTX2_4 were never reported in previous studies. We fit a linear regression model for age prediction based on methylation assay for 89 blood samples from donors aged 9–75 years old. The model included four AR-CpG markers in three gene fragments ASPA, ITGA2B and NPTX2 and enabled the age prediction with R2 = 0.819. The mean absolute deviation (MAD) from chronological age of the model was 7.870. We validated the linear regression model with a validation set of 40 blood samples, and the prediction MAD was 7.986. There was no statistically significant difference in age prediction between 20 pairs of blood samples and bloodstains. Six pairs of fresh and old bloodstains were analyzed using our assay. The obtained results showed the assay still performed an effective prediction on bloodstains after four-month storage in room conditions. This study indicates that our DNA methylation assay is a reliable and effective method for age prediction for forensic purposes. |
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| Keywords: | DNA methylation Age prediction Forensic science Pyrosequencing Human bloodstain Human blood |
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