Abstract
Identification of semen and spermatozoa is crucial in the forensic investigation of alleged sexual assault cases. In cases of alleged sexual assault where there is a long time gap between the incident and sample collection, or in cases of low sperm count, current methods have limitations of specificity, in the case of presumptive tests for semen, or the problem of recording spermatozoa by microscopy if they are few in number. A 3-plex MSRE-PCR (methylation-sensitive restriction enzyme-PCR) assay using a spermatozoa-specific DNA methylated marker to identify spermatozoa has been reported previously by our laboratory. A key advantage over current methods is the increased sensitivity and specificity. A transition from a research tool to operational use requires blind trial testing and inter-laboratory trials. We report on a collaborative exercise where reagents of the 3-plex MSRE-PCR were sent to six participating laboratories. Each laboratory used their own equipment, consumables, and the presumptive reagents conventionally for body fluid (such as acid phosphatase or PSA), DNA extraction, and quantification in practical casework. The reagents and protocol for the 3-plex MSRE-PCR assay and 9 samples were provided by the organizing laboratory. The participating laboratories were requested to fill in the questionnaire after testing. The reported results from all the six participating laboratories were concordant and the expected correct results for the presence of spermatozoa. These outcomes verified the reproducibility and feasibility of the 3-plex MSRE-PCR assay. The results also indicated that the 3-plex MSRE-PCR assay was readily accessible to forensic laboratories for integrating it into current forensic casework processes.
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Acknowledgements
We thank the Ministry of Science and Technology, and Ministry of the Interior of Taiwan who supported the study by the grants numbering NSC 102-2628-B-015-001-MY2 and 108-0805-05-17-01/109-0805-05-17-01, respectively.
Funding
This study was supported by the Ministry of Science and Technology (NSC 102-2628-B-015-001-MY2), and Ministry of the Interior (108-0805-05-17-01 and 109-0805-05-17-01) in Taiwan.
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Material preparation, data collection, and analysis were performed by Kuo-Lan Liu, Li-Chin Tsai, and Yu-Hsuan Chang. The tests for the collaborative exercise were performed by Yu-Chih Lin, Nu-En Huang, Fang-Chin Wu, Chun-Yen Lin, Kuo-Cheng Huang, Chiang-Ho Chen, and Tung-Ho Hsieh. The first draft of the manuscript was written by Lih-Jing Yang, James Chun-I Lee, Adrian Linacre, and Hsing-Mei Hsieh, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
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In this study, samples were collected after informed consent and following the procedures approved by the Institutional Review Board (IRB) of Antai-Tian-Sheng Memorial Hospital (IRB No. 18-074-B) in Taiwan. None of the authors is affiliated to the hospital and the study was not carried out at the hospital. This study was performed in accordance with the ethical standards as laid down in the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards.
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Informed consent was obtained from all individual participants included in the study.
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The authors declare no competing interests.
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Liu, KL., Tsai, LC., Lin, YC. et al. Spermatozoa identification by the 3-plex MSRE-PCR assay: a collaborative exercise. Int J Legal Med 136, 397–404 (2022). https://doi.org/10.1007/s00414-021-02737-2
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DOI: https://doi.org/10.1007/s00414-021-02737-2