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Updated: Nov 27, 2025

Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing
Published on: February 10, 2023
A minimal number CpGs of ELOVL2 gene for a chronological age estimation using pyrosequencing
Poonyapat Sukawutthiya1, Tikumphorn Sathirapatya1, Kornkiat Vongpaisarnsin2
1Department of Forensic Medicine, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand; Forensic Genetics Research Unit, Ratchadapiseksompotch Fund, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.
Abstract:
Chronological age estimation is an important piece of human identification used in forensic practice. Epigenetic modifications, especially DNA methylation, have been proposed to predict age. The methylation of the ELOVL2 gene is one of the age-related markers that could be tested in fresh or postmortem blood sample. We study the use of DNA methylation markers on the ELOVL2 gene and develop a prediction model to estimate the age from a postmortem blood sample using pyrosequencing. From 100 anonymous blood samples, a correlation study of DNA methylation and age was investigated. The regression analysis revealed 2 CpG sites for model prediction with an adjusted R2 value of 0.7 (p < 0.01). The model explained 74% of the variation in postmortem blood samples (n = 36) with a prediction error (RMSE) of 10.2 years and a mean absolute deviation (MAD) of 7.1 years, whereas the model (excluding a younger age group) had improved with a RMSE of 5.6 years and a MAD of 4.2 years. The performance parameters were analyzed in several simulated models and indicated that these markers are advantageous for age estimation in forensic scenarios. Finally, a robustness and reproducibility of the pyrosequencing technique would enable this approach to be the part of an age prediction in forensic investigation.

