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A novel co-amplification system for simultaneous amplification of 23 Y-STR and identification of spermatozoa

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Abstract

In alleged sexual assault cases, identification of the presence of spermatozoa at the crime scene, or on items of eventual significance, or associated with the body of the victim, is integral to the forensic investigation to support or refute the proposition that sexual act has occurred. A 3-plex MSRE-PCR (methylation-sensitive restriction enzyme-PCR) system has been developed previously to identify spermatozoa based on the presence or absence of DNA methylation. This assay showed that 0.1 ng of DNA from a semen extract was sufficient to identify the presence of spermatozoa even when there was excessively more DNA isolated from vaginal fluid than DNA from a semen extract (80 ng/0.1 ng) or a mix of the menstrual blood/semen DNA (5 ng/0.1 ng). In this study, we combine spermatozoa detection with co-amplification of 23 Y-STR loci. We perform standard validation steps to present a novel test that saves time and uses the same sample for both DNA typing and spermatozoa detection in the same reaction. The combined assay can identify Y-STR and spermatozoa simultaneously using just 0.1 ng semen DNA, even in the presence of 5 ng of DNA from a female (male/female:1/50). No other body fluid tested, such as saliva, gave a result for the presence of spermatozoa. A total of 9 non-probative forensic samples from 7 sexual assault cases were tested by this co-amplification system. In all cases, the same sperm-positive data were obtained, concordant with our previous study analyzed by only 3-plex MSRE-PCR, and the Y-STR results were also consistent with that analyzed by only PowerPlex® Y23 kit. The co-amplification will be beneficial for the limited samples in many criminal cases.

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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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Authors and Affiliations

  1. Taichung City Government Police Department, No.500 Fengxing Road Section 1, Tanzi District, Taichung City, 427003, Taiwan, Republic of China

    Yu-Chih Lin

  2. Department of Forensic Science, Central Police University, 56 Shu-Jen Road, Kwei-San, Taoyuan, 33304, Taiwan, Republic of China

    Li-Chin Tsai & Hsing-Mei Hsieh

  3. Forensic Examination Division, Criminal Investigation Bureau, National Police Agency, No.5 Lane 553, Chung Hsiao East Road Section 4, Xinyi District, Taipei, 11072, Taiwan, Republic of China

    Kuo-Lan Liu

  4. Forensic Biology Division, Criminal Investigation Bureau, National Police Agency, No.5 Lane 553, Chung Hsiao East Road Section 4, Xinyi District, Taipei, 11072, Taiwan, Republic of China

    Nu-En Huang & Chih-Wen Su

  5. International Criminal Affairs Division, Criminal Investigation Bureau, National Police Agency, No.5 Lane 553, Chung Hsiao East Road Section 4, Xinyi District, Taipei, 11072, Taiwan, Republic of China

    Lih-Jing Yang

  6. Department of Forensic Medicine, College of Medicine, National Taiwan University, 1 Jen-Ai Road Section 1, Taipei, 10051, Taiwan, Republic of China

    James Chun-I Lee

  7. College of Science & Engineering, Flinders University, Adelaide, 5001, Australia

    Adrian Linacre

Authors
  1. Yu-Chih Lin
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  2. Li-Chin Tsai
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  3. Kuo-Lan Liu
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  4. Nu-En Huang
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  5. Lih-Jing Yang
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  6. Chih-Wen Su
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  7. James Chun-I Lee
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  8. Adrian Linacre
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  9. Hsing-Mei Hsieh
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Contributions

All authors contributed to the conception of this study and the experimental design. Material preparation, data collection, and analyses were performed by Yu-Chih Lin, Li-Chin Tsai, Kuo-Lan Liu, Nu-En Huang, and Chih-Wen Su. 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 subsequent versions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Hsing-Mei Hsieh.

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Ethical approval

In this study, samples were collected after informed consent and following the procedures approved by the Institutional Review Board (IRB) of Tao-Yuan General Hospitals (IRB No. TYGH102011) and Antai-Tian-Sheng Memorial Hospital (IRB No. 18–074-B) in Taiwan. Neither the authors are affiliated to these hospitals nor the study was carried out in these hospitals. And the studies have been 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.

Conflict of interest

The authors declare no competing interests.

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Lin, YC., Tsai, LC., Liu, KL. et al. A novel co-amplification system for simultaneous amplification of 23 Y-STR and identification of spermatozoa. Int J Legal Med 136, 73–84 (2022). https://doi.org/10.1007/s00414-021-02723-8

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  • DOI: https://doi.org/10.1007/s00414-021-02723-8

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