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

Shanan S. Tobe, Yuvaneswari C. Swaran, Lynn Dennany, Ursula Sibbing, Kristina Schulze Johann, Lindsey Welch, Marielle Vennamann

Research output: Contribution to journalArticlepeer-review

8 Citations (Scopus)
150 Downloads (Pure)

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. (150)
Original languageEnglish
Pages (from-to)87-94
Number of pages8
JournalInternational Journal of Legal Medicine
Volume131
Early online date10 Nov 2016
DOIs
Publication statusPublished - 31 Jan 2017

Keywords

  • direct PCR
  • sexual assault
  • semen
  • seminal fluid
  • differential isolation
  • spermatozoa

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