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Bioluminescent assay of microbial ATP in postmortem tissues for the estimation of postmortem interval
1Faculty of Forensic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China. caixe_liu0222@hotmail.com
Abstract:
To study the relationship between changes of microbial ATP in four kinds of murine tissues and the postmortem interval (PMI), healthy SD rats were sacrificed and their muscles, livers, spleens and kidneys were sampled at different postmortem intervals. The concentration of microbial ATP was detected using bioluminescent assay and the data was statistically analyzed. The concentration of microbial ATP in muscle increased with PMI time. The peak appeared at the 7th day after death, and at the 10th day, microbial ATP in muscle tissue increased again. In internal organs, the peaks of microbial ATP were observed at the 8th day after death and the level decreased during 8-10 d. The differences in microbial ATP concentration in liver, spleen and kidney were not statistically significant. During day 0 to day 9 after death, the correlation was best between PMI and microbial ATP in muscle. With PMI as the independent variable, the cubic polynomial regression equation was Y=0.02X(3)-0.166X(2)-0.666X+13.412 (R (2)=0.989, P<0.01). In internal organs, the best correlation was found between PMI and microbial ATP during day 0 to day 10. With PMI as the independent variable, the cubic polynomial regression equation was Y=0.016X(3)-0.127X(2)-0.809X+13.324 (R (2)=0.986, P<0.01). There existed high correlations between PMI and microbial ATP concentration in rat tissues. Since only a small amount of tissue was needed for the detection and the sample was not affected by self-decomposition, the method may extend the time range of PMI estimation.
Insights
Microbial ATP levels in rat tissues correlate with postmortem interval (PMI). Muscle tissue showed the strongest correlation, offering a reliable method for estimating PMI even with small samples.
Area of Science:
- Forensic Science
- Biochemistry
- Microbiology
Background:
- Accurate postmortem interval (PMI) estimation is crucial in forensic investigations.
- Traditional methods for PMI determination have limitations, especially in later stages.
- Microbial ATP as a biomarker for PMI is an emerging area of research.
Purpose of the Study:
- To investigate the relationship between microbial adenosine triphosphate (ATP) concentrations in different murine tissues and the postmortem interval (PMI).
- To establish a reliable method for estimating PMI using microbial ATP levels in tissues.
Main Methods:
- Healthy Sprague-Dawley rats were used, and muscle, liver, spleen, and kidney tissues were collected at various postmortem intervals.
- Microbial ATP concentrations were quantified using bioluminescent assays.
- Statistical analysis and cubic polynomial regression were employed to determine correlations between microbial ATP and PMI.
Main Results:
- Microbial ATP concentration in muscle tissue generally increased with PMI, with notable peaks.
- Internal organs showed peak microbial ATP levels around 8 days postmortem, with subsequent decreases.
- High correlations (R² > 0.98) were found between PMI and microbial ATP in both muscle and internal organs, with specific regression equations derived.
Conclusions:
- Microbial ATP concentration in rat tissues demonstrates a strong correlation with PMI.
- This method requires only small tissue samples and is not significantly affected by autolysis, potentially extending the reliable range for PMI estimation.
- Microbial ATP analysis offers a promising tool for forensic science in determining PMI.
