Application of the Methylated Markers (Spectrin Beta and DEAD-Box Protein) for Definitive Differentiation Between Fresh and Aged Semen by evaluating Their Role in Identifying Semen From Mixed Body Fluids
International Journal of Medical Toxicology and Forensic Medicine,
Vol. 12 No. 4 (2022),
Background: Semen identification is assumed a crucial proof of sexual assault. Moreover, body fluids at the crime scene of a human being, such as blood, semen, and saliva, are often mixed.
Methods: Hence, in our study, we aimed to use methylation analysis targeting DNA epigenetic markers Spectrin beta chain (B_SPTB_03) and DEAD-box protein (DDX4) to differentiate between fresh semen (less than 4 hours) and aged semen (after 24 hours) as well as to differentiate between semen alone and semen mixed with other body fluids (blood and saliva) in the fresh and dried state.
Results: Our findings showed statistically significant differences in the methylation patterns of the SPTB and DDX4 loci to distinguish semen from mixed body fluids in fresh and old samples. We were able to obtain two novel cutoff values to differentiate between fresh and aged semen, which are (52.25) with the SPTB marker and (70.75) with the DDX4 marker.
Conclusion: It is concluded that the methylation approach based on the epigenetic markers of Spectrin beta chain and DEAD-box protein (B_SPTB_03 and DDX4) successfully identified fresh from aged semen and semen-derived alleles from mixed stains, hence it is recommended to be employed in forensic practice.
- Body fluid identification
- DNA methylation
- Forensic application
- Spectrin beta (SPTB)
- DEAD-box protein (DDX4)
How to Cite
Wasserstrom A, Frumkin D, Davidson A, Shpitzen M, Herman Y, Gafny R. Demonstration of dsi-semen--a novel DNA methylation-based forensic semen identification assay. Forensic Science International: Genetics. 2013; 7(1):136-42. [DOI:10.1016/j.fsigen.2012.08.009] [PMID]
An JH, Shin KJ, Yang WI, Lee HY. Body fluid identification in forensics. BMB Reports. 2012; 45(10):545-53.[DOI:10.5483/BMBRep.2012.45.10.206] [PMID]
Pood E. Methylation-specific differentiation of vaginal epithelial cells for forensic tissue typing by bisulfite conversion and pyrosequencing [MSc Thesis]. Hattiesburg: University of Southern Mississippi; 2019. [Link]
Sijen T. Molecular approaches for forensic cell type identification: On mRNA, miRNA, DNA methylation and microbial markers. Forensic Science International: Genetics. 2015; 18:21-32. [DOI:10.1016/j.fsigen.2014.11.015] [PMID]
Balamurugan K, Bombardi R, Duncan G, McCord B. Identification of spermatozoa by tissue-specific differential DNA methylation using bisulfite modification and pyrosequencing. Electrophoresis. 2014; 35(21-22):3079-86. [DOI:10.1002/elps.201400175] [PMID]
Park JL, Kwon OH, Kim JH, Yoo HS, Lee HC, Woo KM, et al. Identification of body fluid-specific DNA methylation markers for use in forensic science. Forensic Science International: Genetics. 2014; 13:147-53. [DOI:10.1016/j.fsigen.2014.07.011] [PMID]
Haas C, Klesser B, Maake C, Bär W, Kratzer A. mRNA profiling for body fluid identification by reverse transcription endpoint PCR and realtime PCR. Forensic Science International: Genetics. 2009; 3(2):80-8. [DOI:10.1016/j.fsigen.2008.11.003] [PMID]
Watanabe K, Akutsu T, Takamura A, Sakurada K. Evaluation of a blood-specific DNA methylated region and trial for allele-specific blood identification from mixed body fluid DNA. Legal Medicine. 2016; 22:49-53. [DOI:10.1016/j.legalmed.2016.08.004] [PMID]
Lee HY, Park MJ, Choi A, An JH, Yang WI, Shin KJ. Potential forensic application of DNA methylation profiling to body fluid identification. International Journal of Legal Medicine. 2012; 126(1):55-62. [DOI:10.1007/s00414-011-0569-2] [PMID]
Brait M, Maldonado L, Begum S, Loyo M, Wehle D, Tavora FF, et al. DNA methylation profiles delineate epigenetic heterogeneity in seminoma and non-seminoma. British Journal of Cancer. 2012; 106(2):414-23. [DOI:10.1038/bjc.2011.468] [PMID] [PMCID]
Pfaffl M. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Research. 2001; 29(9):e45. [DOI:10.1093/nar/29.9.e45] [PMID] [PMCID]
Chan YH. Biostatistics 102: Quantitative data--parametric & non-parametric tests. Singapore Medical Journal. 2003; 44(8):391-6. [PMID]
Ledgerwood SM. High resolution melt analysis of samples with differential DNA methylation to identify tissue source of origin [MSc Thesis]. Hattiesburg: University of Southern Mississippi; 2015. [Link]
Blackman S, Stafford-Allen B, Hanson EK, Panasiuk M, Brooker AL, Rendell P, et al. Developmental validation of the paraDNA® body fluid id system-a rapid multiplex mRNA-profiling system for the forensic identification of body fluids. Forensic Science International: Genetics. 2018; 37:151-61. [DOI:10.1016/j.fsigen.2018.08.012] [PMID]
Stravers CS, Gool EL, van Leeuwen TG, Aalders MC, van Dam A. Multiplex body fluid identification using surface plasmon resonance imaging with principal component analysis. Sensors and Actuators B: Chemical. 2019; 283:355-62. [DOI:10.1016/j.snb.2018.12.050]
Abbas N, Lu X, Badshah M, il Heo W, Seo SJ, Kim SM. Identification of body fluids using metal enhanced fluorescence substrate with glancing angle deposited Ag nanorods. Paper presented at: Frontiers in Biological Detection: From Nanosensors to Systems XI. 4 March 2019; San Francisco: United States. [DOI:10.1117/12.2511045] [PMCID]
Zapata F, de la Ossa MÁ, García-Ruiz C. Differentiation of body fluid stains on fabrics using external reflection fourier transform infrared spectroscopy (FT-IR) and chemometrics. Applied Spectroscopy. 2016; 70(4):654-65. [DOI:10.1177/0003702816631303] [PMID]
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