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A proof of principal study on the use of direct PCR of semen and spermatozoa and development of a differential isolation protocol for use in cases of alleged sexual assault

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Abstract

Sexual assault samples are some of the most common samples encountered in forensic analysis. These samples can require a significant time investment due to differential extraction processes. We report on the first record of successful direct amplification of semen for STR analysis. Neat seminal fluid, dilutions ranging from 1:5 to 1:160 and GEDNAP samples were successfully amplified using a direct method. A mild differential isolation technique to enrich spermatozoa was developed and successfully implemented to separate and directly amplify a mixture of semen and female epithelial cells. Aliquots of samples subjected to the differential isolation protocol were stained with Haemotoxylin and Eosin for sperm scoring. Samples stained after PCR showed a complete lack of intact spermatozoa demonstrating that the cells are lysed during the PCR process. This paper demonstrates the potential to incorporate direct PCR in cases of sexual assault to more rapidly obtain results and achieve a higher sensitivity.

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Acknowledgments

YCS was funded through the Government of Malaysia. We would also like to thank Lisa Dierig and Marianne Schürenkamp for the excellent technical assistance and Prof. Peter Schmidt (Münster) for the help with microscopic images.

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Author notes

  1. Yuvaneswari C. Swaran

    Present address: Forensic Division, Department of Chemistry Malaysia, Jalan Sultan, Malaysia

Authors and Affiliations

  1. Department of Chemistry and Physics, Arcadia University, 450 S. Easton Rd., Glenside, PA, 19038, USA

    Shanan S. Tobe

  2. School of Biological Sciences, Flinders University, Adelaide, Australia

    Shanan S. Tobe

  3. Centre for Forensic Science, WestCHEM, Technology and Innovation Centre, University of Strathclyde, Glasgow, UK

    Yuvaneswari C. Swaran, Lynn Dennany & Lindsey Welch

  4. Institute of Legal Medicine, University of Münster, Münster, Germany

    Ursula Sibbing, Kristina Schulze Johann & Marielle Vennemann

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  1. Shanan S. Tobe
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  2. Yuvaneswari C. Swaran
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  3. Lynn Dennany
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Correspondence to Shanan S. Tobe.

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Tobe, S.S., Swaran, Y.C., Dennany, L. et al. A proof of principal study on the use of direct PCR of semen and spermatozoa and development of a differential isolation protocol for use in cases of alleged sexual assault. Int J Legal Med 131, 87–94 (2017). https://doi.org/10.1007/s00414-016-1461-x

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  • DOI: https://doi.org/10.1007/s00414-016-1461-x

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