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. 2025 Jan;139(1):175-185.
doi: 10.1007/s00414-024-03320-1. Epub 2024 Sep 13.

Age estimation of burnt human remains through DNA methylation analysis

Pierangela Grignani  1 , Barbara Bertoglio  2 , Maria Cristina Monti  1 , Riccardo Cuoghi Costantini  3 , Ugo Ricci  4 , Martina Onofri  5 , Paolo Fattorini  6 , Carlo Previderè  1
Affiliations

Age estimation of burnt human remains through DNA methylation analysis

Pierangela Grignani et al. Int J Legal Med. 2025 Jan.

Abstract

The identification of human fire victims is a challenging task in forensic medicine. The heat-induced alterations of biological tissues can make the conventional anthropological analyses difficult. Even if the DNA profile of the victim is achieved, it is possible that no match can be found in a forensic DNA database, thus hindering positive identification. In such cases, any information useful to nail down a possible identity should be collected, such as DNA methylation analysis which could provide useful investigative leads. In the present study, five age-related epigenetic markers (ELOVL2, FHL2, KLF14, C1orf132, and TRIM59) were initially analysed in blood samples of 72 living Italian individuals of known age, using a Single Base Extension (SBE) assay. An age prediction model was built by multiple linear regression including all the markers (Mean Absolute Error, MAE: 3.15 years). This model was tested on 29 blood samples collected during autopsies from burnt human remains, already identified through DNA analysis, providing a MAE of 6.92 years. The model allowed a correct prediction in 79.3% of the cases (95% prediction interval), while six cases were associated with inaccurate predictions (min-max prediction error: 9.8-37.3 years). Among the different sample variables considered to explain these results, only the DNA degradation index was a relevant factor affecting the reliability of the predictions. In conclusion, the SBE typing of blood from burnt remains proved to be a reliable tool to estimate chronological age of most of the samples, also in consideration of its cost-effectiveness and the availability of CE sequencers in every forensic genetics laboratory.

Keywords: Age estimation; Burnt remains; DNA methylation; Single base extension (SBE).

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Conflict of interest statement

Declarations. Ethics approval: This study was performed in line with the principles of the Declaration of Helsinki. Approval was granted by the Ethic Committee of the University of Trieste (Comitato Etico di Ateneo, number 135, October 24th, 2023). Consent to participate: Informed consent was obtained from all individual participants included in the study. Competing interests: The authors have no conflict of interest to declare.

Figures

Fig. 1
Fig. 1
Forest plot representing the estimated regression model coefficients (β) and 95% confidence intervals with the corresponding t-statistic (t) and associated p-value (p) explaining the statistical significance of each model term
Fig. 2
Fig. 2
Prediction error versus chronological age for the living (a) and burnt human remains (b). Correct and not correct predictions are shown in black and red, respectively
See this image and copyright information in PMC

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