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Construction and validation of a rapid semen identification system based on SHERLOCK technology
Yu Luo1, Xianmiao Wang2, Fan Yang3
1Department of Basic Medical Sciences, Qinghai Unversity Medical College, Xining, Qinghai 810016, China; Key Laboratory of Forensic Genetics of Ministry of Public Security, Institute of Forensic Science of China, Beijing 100038, China.
Forensic Science International. Genetics
|December 12, 2025
Summary
Forensic scientists can now rapidly identify semen samples using a new CRISPR/Cas13a-based detection system. This innovative method offers high sensitivity and specificity for on-site body fluid identification.
Area of Science:
- Forensic Science
- Molecular Biology
- Biotechnology
Background:
- Accurate and timely identification of biological evidence is crucial in forensic investigations.
- Existing methods for body fluid identification can be time-consuming and require specialized laboratory equipment.
- There is a need for rapid, on-site detection methods for forensic applications.
Purpose of the Study:
- To develop a rapid, sensitive, and specific detection system for semen-specific mRNA for forensic use.
- To integrate CRISPR/Cas13a technology with rapid sample preparation for on-site application.
- To evaluate the performance of the developed method in terms of specificity and sensitivity.
Main Methods:
- Design and screening of specific primers and CRISPR RNA (crRNA) targeting semen-specific mRNA.
- Establishment of a SHERLOCK (Specific High-sensitivity Enzymatic Reporter unLOCKing) detection assay based on CRISPR/Cas principles.
- Screening of nucleic acid rapid release agents for efficient sample treatment.
- Integration of rapid extraction with the SHERLOCK assay for a streamlined detection process.
Main Results:
- The developed method successfully detected semen-specific mRNA with high specificity.
- The system demonstrated a sample detection sensitivity as low as 0.25 μL of semen.
- Semen samples showed significantly higher relative fluorescence unit (RFU) values compared to non-semen samples (P < 0.0001).
Conclusions:
- The novel CRISPR/Cas13a-based system provides a rapid and reliable method for on-site semen detection.
- This approach significantly reduces operational time and dependence on complex instrumentation.
- The developed method offers an innovative solution for real-time forensic body fluid identification.
