<- Home <- Arhive <- Vol. 29, Issue 1, March 2021



Rom J Leg Med29(1)1-7(2021)
DOI:10.4323/rjlm.2021.1
© Romanian Society of Legal Medicine


ROLE OF CELL FREE DNA AND HMGB-1 IN POSTMORTEM INTERVAL DETERMINATION

E. Emiral, D. Y. Kilicaslan, I. H. Hanci, N. L. S. Tufan,


Abstract: Postmortem interval (PMI) determination is essential for criminal and civil cases concerning forensic death. Today, many methods that are in practice are subjective, without a conclusive result. With the increasing use of molecular studies in forensic sciences, key studies have been conducted especially in the field of Forensic Genetics. This study aimed to determine PMI by utilizing early postmortem changes in cell free DNA and serum HMGB-1 protein levels that particularly increase in serum with cell necrosis.
The study was carried out on 96 Wistar rats, weighing 230–260 g. The rats were kept at two different temperatures, i.e. +4°C and +24°C, after anesthesia and cervical dislocation. Blood samples were collected during autopsy at hours 0, 3, 6, 9, 12, 24, 48 and 72. The blood was centrifuged for five minutes at 5000 rpm to separate the serum. HMGB-1 concentration measurement in serum samples using HMGB-1 ELISA kit and SYBR Gold Nucleic Acid Gel Stain nucleic acid staining protocol was followed by optical density measurement with a luminometer for cell free DNA.
The results obtained in the study were converted into concentration values by equations obtained from standard samples. It was found that there was an increase in cell free DNA and HMGB-1 level in serum in parallel to the increasing length postmortem interval at +4°C ([r = 0.751 p < 0.001] and [r = 0.698 p < 0.001], respectively). At +24°C, postmortem interval was found to have a negative correlation with the amount of cell free DNA in serum (r = −0.213 p = 0.15) and a weak positive correlation with serum HMGB-1 concentration (r = 0.313 p = 0.030).
In conclusion, our study was the first experimental study to investigate cell free DNA and serum HMGB-1 levels together in postmortem interval determination. Our findings suggest that serum cell free DNA and serum HMGB-1 levels at +4°C, where decay is particularly less effective, can be used in the determination of PMI.
Keywords: Postmortem interval, HMGB-1, Cell Free DNA, Time of death, ELISA



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