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Related Experiment Video

Updated: May 21, 2026

Methylation Specific Multiplex Droplet PCR using Polymer Droplet Generator Device for Hematological Diagnostics
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Methylation Specific Multiplex Droplet PCR using Polymer Droplet Generator Device for Hematological Diagnostics

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DNA methylation-specific multiplex assays for body fluid identification.

Ja Hyun An1, Ajin Choi, Kyoung-Jin Shin

  • 1Department of Forensic Medicine and Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine, 50 Yonsei-ro, Seodaemun-gu, Seoul, 120-752, South Korea.

International Journal of Legal Medicine
|June 2, 2012
PubMed
Summary

DNA methylation profiles in semen-specific regions are stable over time and can identify semen, differentiating it from other body fluids. This epigenetic approach shows promise for forensic body fluid identification.

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Corrigendum to "DNA methylation profiling for a confirmatory test for blood, saliva, semen, vaginal fluid and menstrual blood" [Forensic Sci. Int. Genet. 24 (2016) 75-82].

Forensic science international. Genetics·2026
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COSA: A consolidated methylation-based prediction panel for body fluid/tissue of origin, smoking status, and chronological age using amplicon-based massively parallel sequencing.

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Erratum to "Age prediction using DNA methylation of Y-chromosomal CpGs in semen samples" [Forensic Sci. Int.: Genet. 69 (2024) 103007].

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Y-CpG-based semen age prediction: analysis of vasectomized samples and development of an optimized multiplex assay evaluated in independent and mixed samples.

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A strategy for body fluid mixture analysis using semen- and epidermis-specific tDMRs and adjacent microhaplotypes/SNPs.

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Enhancing forensic casework interpretation through DNA methylation-based analysis: A case study of pooled blood samples.

Forensic science international. Genetics·2026

Area of Science:

  • Forensic science
  • Epigenetics
  • Molecular biology

Background:

  • Tissue-specific differentially methylated regions (tDMRs) exhibit unique DNA methylation patterns across different cell and tissue types.
  • Differential DNA methylation in body fluids offers a potential method for identification.
  • DNA methylation can be influenced by environmental factors and aging, necessitating investigation into its stability for forensic applications.

Purpose of the Study:

  • To investigate age-related methylation changes in semen-specific tDMRs.
  • To assess the stability of body fluid-specific DNA methylation profiles over time.
  • To develop and validate multiplex PCR systems for body fluid identification using DNA methylation analysis.

Main Methods:

  • Analysis of age-related methylation changes in semen-specific tDMRs from young and elderly men.

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  • Confirmation of the stability of body fluid-specific DNA methylation profiles.
  • Development of two multiplex PCR systems (methylation-sensitive restriction enzyme PCR and methylation SNaPshot) to analyze CpG site methylation status in USP49, DACT1, PRMT2, and PFN3 tDMRs.

    • Main Results:

    • The study confirmed the stability of body fluid-specific DNA methylation profiles over time.
    • Both developed multiplex PCR systems successfully identified semen containing spermatozoa.
    • The systems could differentiate menstrual blood and vaginal fluids from blood and saliva.

    Conclusions:

    • DNA methylation profiling is a potentially valuable tool for forensic body fluid identification.
    • While semen-specific tDMRs show stable methylation patterns, further markers may enhance identification accuracy.
    • The developed multiplex PCR systems demonstrate the feasibility of using epigenetic markers for forensic analysis of body fluids.