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Related Experiment Video

Updated: May 15, 2025

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions
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Development of a Codetection System Based on STRs and Biofluid-Specific CpG Markers.

Zeqin Li1,2, Shiqi Liu1,3, Fang Yuan1,2

  • 1School of Forensic Medicine, Shanxi Medical University, Jinzhong, Shanxi, China.

Biotechnology and Applied Biochemistry
|April 8, 2025
PubMed
Summary

Forensic scientists can now simultaneously identify body fluids and their donors using a novel DNA analysis system. This integrated approach enhances evidence validity in criminal investigations by combining methylation markers and short tandem repeats (STR) analysis.

Keywords:
body fluid identificationforensic genetics personal identificationmethylation‐sensitive restriction enzyme‐polymerase chain reaction (MSRE‐PCR)short tandem repeats

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Area of Science:

  • Forensic Science
  • Molecular Biology
  • Genetics

Background:

  • Accurate identification of body fluids and their donors is crucial for forensic investigations.
  • Existing methods for personal identification (autosomal short tandem repeats) and body fluid origin (DNA methylation) are often performed separately.
  • There is a need for an integrated system for simultaneous subsource and source-level analysis.

Purpose of the Study:

  • To develop and validate a codetection system for simultaneous personal identification and biofluid origin determination.
  • To integrate DNA methylation markers with a short tandem repeat (STR) amplification system for capillary electrophoresis.

Main Methods:

  • Developed a multiplex system combining a 7-plex DNA methylation assay (5 biofluid-specific CpG markers and 2 controls) with a 20-plex STR amplification system.
  • Utilized capillary electrophoresis for simultaneous analysis of methylation profiles and STR typing.
  • Tested the system on single-source and mixed body fluid samples (peripheral blood, semen, saliva, menstrual blood, vaginal secretion).

Main Results:

  • The system successfully distinguished five types of body fluids from single sources.
  • Generated DNA methylation profiles for mixed samples containing two to four body fluids in various proportions.
  • Achieved a high cumulative discrimination power for personal identification (1-3.5120 × 10-25).
  • Demonstrated efficient identification of semen and vaginal secretion with low DNA input (0.1 ng) and after prolonged storage (5 months at room temperature).

Conclusions:

  • The developed codetection system offers a powerful tool for simultaneous body fluid identification and donor profiling in forensic science.
  • This integrated approach enhances the efficiency and validity of evidence analysis in criminal investigations.
  • The system shows promise for routine forensic application due to its sensitivity and robustness.