The Effect of Temperature on Age Estimation of Semen Stains on Porous Versus Non-porous Surfaces Using Messenger Ribonucleic Acid Measurement
International Journal of Medical Toxicology and Forensic Medicine,
Vol. 12 No. 4 (2022),
Background: While messenger Ribonucleic Acid (mRNA) can be used to identify the type of body fluid, its degradation can also indicate the time interval since it was deposited. This study was conducted to evaluate the effect of temperature on the estimation of the age of human semen stains using mRNA deposited on porous versus non-porous surfaces at different time intervals.
Methods: Ten semen samples were applied on two different media (glass and cotton) and exposed to three different temperatures (4°C, room temperature, 40°C) and examined at three-time intervals (0, 45, and 90 days). The semen-specific mRNA markers protamine 1 (PRM1) and protamine 2 (PRM2) were quantitatively assessed along with a reference gene, beta-actin, using a reverse transcription-quantitative polymerase chain reaction.
Results: Mean Cq values of mRNA markers (PRM1 and PRM2) and the reference gene (beta-actin) increased with time of storage at different temperatures in both examined media. The mean quantification cycle (Cq) values of PRM2 were lower than PRM1, indicating that the levels of PRM2 marker in semen stain were higher than those of PRM1 marker. However, the mean Cq values of PRM2 at each time interval were not significantly different between temperatures, while PRM1 showed statistically significant differences in mean Cq values between temperatures at day 45 in both media.
Conclusion: These results indicate that PRM2 can act as a reliable mRNA marker to estimate the time of deposition of semen stain at different temperatures on two different media.
- Semen age
- Messenger ribonucleic acid
- Reverse transcription-quantitative polymerase chain reaction
How to Cite
Morling N, Mosquera-Miguel A, -Parson W, Phillips C, Porto MJ, Pośpiech E, et al. Body fluid identification using a targeted mRNA massively parallel sequencing approach - results of a euroforgen/ednap collaborative exercise. Forensic Science International. Genetics. 2018; 34:105-15. [DOI:10.1016/j.fsigen.2018.01.002] [PMID]
Alshehhi S, Haddrill PR. Estimating time since deposition using quantification of RNA degradation in body fluid-specific markers. Forensic Science International. 2019; 298:58-63. [DOI:10.1016/j.forsciint.2019.02.046] [PMID]
Taha E, Gomaa R, Nader L. Validation of recently discovered mrna stable regions as biomarkers for body fluids after exposure to environmental hazards. International Journal of Sciences: Basic and Applied Research (IJSBAR). 2017; 36(5):193-207. [Link]
Zha S, Wei X, Fang R, Wang Q, Lin H, Zhang K, et al. Estimation of the age of human semen stains by attenuated total reflection fourier transform infrared spectroscopy: A preliminary study. Forensic Sciences Research. 2019; 5(2):119-25. [DOI:10.1080/20961790.2019.1642567] [PMID] [PMCID]
Chan YH. Biostatistics102: Quantitative data - parametric & non-parametric tests. Singapore Medical Journal. 2003; 44(8):391-6. [PMID]
Chan YH. Biostatistics 104: Correlational analysis. Singapore Medical Journal. 2003; 44(12):614-19. [PMID]
Heneghan N, Fu J, Pritchard J, Payton M, Allen RW. The effect of environmental conditions on the rate of RNA degradation in dried blood stains. Forensic Science International: Genetics. 2021; 51:102456. [DOI:10.1016/j.fsigen.2020.102456] [PMID]
Schrader C, Schielke A, Ellerbroek L, Johne R. PCR inhibitors - occurrence, properties and removal. Journal of Applied Microbiology. 2012; 113(5):1014-26. [DOI:10.1111/j.1365-2672.2012.05384.x] [PMID]
Sakurada K, Akutsu T, Watanabe K, Miyasaka S, Kasai K. Identification of body fluid stains using real-time RT-PCR: Discrimination between salivary, nasal, and vaginal secretions. Japanese Journal of Forensic Science and Technology. 2013; 18(1):1-1. [DOI:10.3408/jafst.18.1]
Alshehhi S, McCallum NA, Haddrill PR. Quantification of RNA degradation of blood-specific markers to indicate the age of bloodstains. Forensic Science International: Genetics Supplement Series. 2017; 6:e453-5. [DOI:10.1016/j.fsigss.2017.09.175]
Weinbrecht KD, Fu J, Payton M, Allen RW. Time-dependent loss of mRNA transcripts from forensic stains. Research and Reports in Forensic Medical Science. 2017; 7:1-2. [DOI:10.2147/RRFMS.S125782]
Fu J, Allen RW. A method to estimate the age of bloodstains using quantitative PCR. Forensic Science International. Genetics. 2019; 39:103-8. [DOI:10.1016/j.fsigen.2018.12.004] [PMID]
Hermann J, Hoff S, Muñoz-Zanzi C, Yoon KJ, Roof M, Burkhardt A, et al. Effect of temperature and relative humidity on the stability of infectious porcine reproductive and respiratory syndrome virus in aerosols. Veterinary Research. 2007; 38(1):81-93. [DOI:10.1051/vetres:2006044] [PMID]
Sakurada K, Ikegaya H, Fukushima H, Akutsu T, Watanabe K, Yoshino M. Evaluation of mRNA-based approach for identification of saliva and semen. Legal Medicine. 2009; 11(3):125-8. [DOI:10.1016/j.legalmed.2008.10.002] [PMID]
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