Magnetic bead-based separation of sperm cells from semen-vaginal fluid mixed stains using an anti-ACRBP antibody

Int J Legal Med. 2023 Mar;137(2):511-518. doi: 10.1007/s00414-022-02917-8. Epub 2022 Nov 23.

Abstract

Forensic DNA analysis of semen-vaginal fluid mixed stains is essential and necessary in sexual assault cases. Here, we used a magnetic bead conjugated acrosin binding protein (ACRBP) antibody to separate and enrich sperm cells from mixed stains. Previously, western blotting indicated that ACRBP was specifically expressed in sperm cells, but not in female blood and epithelial cells, while immunofluorescence data showed ACRBP was localized to the acrosome in sperm cells. In our study, sperm were separated from mixed samples at three sperm cell/female buccal epithelial cell ratios (103:103; 103:104; and 103:105) using a magnetic bead conjugated ACRBP antibody. Subsequently, 23 autosomal short tandem repeat (STR) loci were amplified using the Huaxia™ Platinum PCR Amplification System and genotyped using capillary electrophoresis. The genotyping success rate for STR loci was 90% when the sperm to female buccal epithelial cell ratio was > 1:100 in mixed samples. Our results suggest that the magnetic bead conjugated ACRBP antibody is effective for isolating sperm cells in sexual assault cases.

Keywords: Acrosin binding protein; Forensic science; Magnetic beads; Mixed stain; Sperm.

MeSH terms

  • Coloring Agents* / metabolism
  • DNA Fingerprinting / methods
  • Female
  • Humans
  • Magnetic Phenomena
  • Male
  • Semen*
  • Spermatozoa
  • Staining and Labeling

Substances

  • Coloring Agents