UNLABELLED: Human age estimation from trace samples may give important leads early in a police investigation by contributing to the description of the perpetrator. Several molecular biomarkers are available for the estimation of chronological age, and currently, DNA methylation patterns are the most promising. In this study, a QIAGEN age protocol for age estimation was tested by five forensic genetic laboratories. The assay comprised bisulfite treatment of the extracted DNA, amplification of five CpG loci (in the genes of ELOVL2, C1orf132, TRIM59, KLF14, and FHL2), and sequencing of the amplicons using the PyroMark Q48 platform. Blood samples from 49 individuals with ages ranging from 18 to 64 years as well as negative and methylation controls were analyzed. An existing age estimation model was applied to display a mean absolute deviation of 3.62 years within the reference data set. KEY POINTS: Age determination as an intelligence tool during investigations can be a powerful tool in forensic genetics.In this study, five laboratories ran 49 samples and obtained a mean absolute deviation of 3.62 years.Five markers were analyzed on a PyroMark Q48 platform.

Department of Chemistry and Biochemistry Florida International University Miami FL USA

Institute of Legal Medicine Faculty of Medicine and University Clinic University of Cologne Cologne Germany

Institute of Molecular and Translational Medicine Faculty of Medicine and Dentistry Palacky University Olomouc and the University Hospital Olomouc Olomouc the Czech Republic

Laboratoire d'Hématologie Médico Légale Bordeaux Cedex France

Section of Forensic Genetics Department of Forensic Medicine Faculty of Health and Medical Sciences University of CopenhagenCopenhagen Denmark

Zobrazit více v PubMed

Aliferi A, Gallidabino MD, Thurtle H, et al. . DNA methylation-based age prediction using massively parallel sequencing data and multiple machine learning models. Forensic Sci Int Genet. 2018;37:215–226. PubMed

Vidaki A, Aliferi A, Miller TH, et al. . DNA methylation-based forensic age prediction using artificial neural networks and next generation sequencing. Forensic Sci Int Genet. 2017;28:225–236. PubMed PMC

Bocklandt S, Lin W, Sehl ME, et al. . Epigenetic predictor of age. PLoS One. 2011;6:e14821. PubMed PMC

Eipel M, Mayer F, Arent T, et al. . Epigenetic age predictions based on buccal swabs are more precise in combination with cell type-specific DNA methylation signatures. Aging (Albany NY). 2016;8:1034–1048. PubMed PMC

Freire-Aradas A, Phillips C, Mosquera-Miguel A, et al. . Development of a methylation marker set for forensic age estimation using analysis of public methylation data and the Agena bioscience EpiTYPER system. Forensic Sci Int Genet. 2016;24:65–74. PubMed

Garagnani P, Bacalini MG, Pirazzini C, et al. . Methylation of ELOVL2 gene as a new epigenetic marker of age. Aging Cell. 2012;11:1132–1134. PubMed

Koch CM, Wagner W. Epigenetic-aging-signature to determine age in different tissues. Aging (Albany NY). 2011;3:1018–1027. PubMed PMC

Park JL, Kim JH, Seo E, et al. . Identification and evaluation of age-correlated DNA methylation markers for forensic use. Forensic Sci Int Genet. 2016;23:64–70. PubMed

Xu C, Qu H, Wang G, et al. . A novel strategy for forensic age prediction by DNA methylation and support vector regression model. Sci Rep. 2015;5:17788. PubMed PMC

Yi SH, Jia YS, Mei K, et al. . Age-related DNA methylation changes for forensic age-prediction. Int J Leg Med. 2015;129:237–244. PubMed

Zbiec-Piekarska R, Spolnicka M, Kupiec T, et al. . Development of a forensically useful age prediction method based on DNA methylation analysis. Forensic Sci Int Genet. 2015;17:173–179. PubMed

Bram Bekaert AK, Zapico SC, Van de Voorde, et al. . Improved age determination of blood and teeth samples using a selected set of DNA methylation markers. Epigenetics. 2015;10:922–930. PubMed PMC

Horvath S. DNA methylation age of human tissues and cell types. Genome Biol. 2013;14:R115. PubMed PMC

Weidner CI, Lin Q, Koch CM, et al. . Aging of blood can be tracked by DNA methylation changes at just three CpG sites. Genome Biol. 2014;15:R24. PubMed PMC

Garcia-Donas JG, Bonicelli A, Scholl AR, et al. . Rib histomorphometry: a reliability and validation study with a critical review of histological techniques for forensic age estimation. Leg Med (Tokyo). 2021;49:101827. PubMed

Ubelaker DH, Khosrowshahi H. Estimation of age in forensic anthropology: historical perspective and recent methodological advances. Forensic Sci Res. 2019;4:1–9. PubMed PMC

Valsecchi A, Irurita Olivares J, Mesejo P. Age estimation in forensic anthropology: methodological considerations about the validation studies of prediction models. Int J Leg Med. 2019;133:1915–1924. PubMed

Ruengdit S, Troy Case D, Mahakkanukrauh P. Cranial suture closure as an age indicator: a review. Forensic Sci Int. 2020;307:110111. PubMed

Hermetet C, Saint-Martin P, Gambier A, et al. . Forensic age estimation using computed tomography of the medial clavicular epiphysis: a systematic review. Int J Leg Med. 2018;132:1415–1425. PubMed

Verma M, Verma N, Sharma R, et al. . Dental age estimation methods in adult dentitions: an overview. J Forensic Dent Sci. 2019;11:57–63. PubMed PMC

Frommer M, Mcdonald LE, Millar DS, et al. . A genomic sequencing protocol that yields a positive display of 5-methylcytosine residues in individual DNA strands. P Natl Acad Sci USA. 1992;89:1827–1831. PubMed PMC

Boyd VL, Zon G. Capillary electrophoretic analysis of methylation status in CpG-rich regions by single-base extension of primers modified with N6-methoxy-2,6-diaminopurine. Anal Biochem. 2008;380:13–20. PubMed

Hong SR, Jung SE, Lee EH, et al. . DNA methylation-based age prediction from saliva: high age predictability by combination of 7 CpG markers. Forensic Sci Int-Gen. 2017;29:118–125. PubMed

Hannum G, Guinney J, Zhang L, et al. . Genome-wide methylation profiles reveal quantitative views of human aging rates. Mol Cell. 2013;49:359–367. PubMed PMC

Nyren P. Enzymatic method for continuous monitoring of DNA polymerase activity. Anal Biochem. 1987;167:235–238. PubMed

Nilsson M, Styrman H, Andréasson H, et al. . Sensitive forensic analysis using the pyrosequencing technology. Int Congr Ser 2006;1288:625–627.

Allen MAH, Andréasson H. Mitochondrial D-loop and coding sequence analysis using pyrosequencing. Methods Mol Biol. 2005;297:179–196. PubMed

Andreasson H, Asp A, Alderborn A, et al. . Mitochondrial sequence analysis for forensic identification using pyrosequencing technology. Biotechniques. 2002;32:124–133. PubMed

Fleckhaus J, Schneider PM. Novel multiplex strategy for DNA methylation-based age prediction from small amounts of DNA via pyrosequencing. Forensic Sci Int Genet. 2020;44:102189. PubMed

Spolnicka M, Pospiech E, Peplonska B, et al. . DNA methylation in ELOVL2 and C1orf132 correctly predicted chronological age of individuals from three disease groups. Int J Leg Med. 2018;132:1–11. PubMed PMC

Dias HC, Cordeiro C, Pereira J, et al. . DNA methylation age estimation in blood samples of living and deceased individuals using a multiplex SNaPshot assay. Forensic Sci Int. 2020;311:110267. PubMed

Freire-Aradas A, Phillips C, Giron-Santamaria L, et al. . Tracking age-correlated DNA methylation markers in the young. Forensic Sci Int Genet. 2018;36:50–59. PubMed