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

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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.

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References 

  1. Tan Y et al (2018) Two-person DNA mixture interpretation based on a novel set of SNP-STR markers. Forensic Sci Int Genet 37:37–45

    Article  CAS  Google Scholar 

  2. Andersen MM et al (2015) Identifying the most likely contributors to a Y-STR mixture using the discrete Laplace method. Forensic Sci Int Genet 15:76–83

    Article  CAS  Google Scholar 

  3. Bieber FR et al (2016) Evaluation of forensic DNA mixture evidence: protocol for evaluation, interpretation, and statistical calculations using the combined probability of inclusion. BMC Genet 17(1):125

    Article  Google Scholar 

  4. Riman S, Iyer H, Vallone PM (2021) Examining performance and likelihood ratios for two likelihood ratio systems using the PROVEDIt dataset. PLoS ONE 16(9):e0256714

    Article  CAS  Google Scholar 

  5. Coble MD, Bright JA (2019) Probabilistic genotyping software: an overview. Forensic Sci Int Genet 38:219–224

    Article  CAS  Google Scholar 

  6. Moretti TR et al (2017) Internal validation of STRmix for the interpretation of single source and mixed DNA profiles. Forensic Sci Int Genet 29:126–144

    Article  CAS  Google Scholar 

  7. Timken MD, Klein SB, Buoncristiani MR (2018) Improving the efficacy of the standard DNA differential extraction method for sexual assault evidence. Forensic Sci Int Genet 34:170–177

    Article  CAS  Google Scholar 

  8. Xu Y et al (2016) Fluorescence- and magnetic-activated cell sorting strategies to separate spermatozoa involving plural contributors from biological mixtures for human identification. Sci Rep 6:36515

    Article  CAS  Google Scholar 

  9. Dean L et al (2015) Separation of uncompromised whole blood mixtures for single source STR profiling using fluorescently-labeled human leukocyte antigen (HLA) probes and fluorescence activated cell sorting (FACS). Forensic Sci Int Genet 17:8–16

    Article  CAS  Google Scholar 

  10. Vandewoestyne M, Deforce D (2010) Laser capture microdissection in forensic research: a review. Int J Legal Med 124(6):513–521

    Article  Google Scholar 

  11. Cerri N et al (2003) Mixed stains from sexual assault cases: autosomal or Y-chromosome short tandem repeats? Croat Med J 44(3):289–292

    Google Scholar 

  12. Andersen MM, Balding DJ (2017) How convincing is a matching Y-chromosome profile? PLoS Genet 13(11):e1007028

    Article  Google Scholar 

  13. Kannourakis G, Bol S (1987) Fractionation of normal and beta-thalassemic human hemopoietic progenitor cells by immunomagnetic beads. Exp Hematol 15(11):1103–1108

    CAS  Google Scholar 

  14. Ravelo KM, Andersen ND, Monje PV (2018) Magnetic-activated cell sorting for the fast and efficient separation of human and rodent Schwann cells from mixed cell populations. Methods Mol Biol 1739:87–109

    Article  CAS  Google Scholar 

  15. Li XB et al (2014) Magnetic bead-based separation of sperm from buccal epithelial cells using a monoclonal antibody against MOSPD3. Int J Legal Med 128(6):905–911

    Article  Google Scholar 

  16. Zhao XC et al (2016) Isolating sperm from cell mixtures using magnetic beads coupled with an anti-PH-20 antibody for forensic DNA analysis. PLoS ONE 11(7):e0159401

    Article  Google Scholar 

  17. Lin L et al (2021) Cancer-testis antigen ACRBP expression and serum immunoreactivity in ovarian cancer: its association with prognosis. Immun Inflamm Dis 9(4):1759–1770

    Article  CAS  Google Scholar 

  18. Tanphaichitr N et al (2015) Remodeling of the plasma membrane in preparation for sperm-egg recognition: roles of acrosomal proteins. Asian J Androl 17(4):574–582

    Article  CAS  Google Scholar 

  19. Walsh PS, Metzger DA, Higushi R (1991) Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material. BioTechniques 10(4):506–13

    CAS  Google Scholar 

  20. Li W et al (2020) Forensic characteristics and phylogenetic analyses of one branch of Tai-Kadai language-speaking Hainan Hlai (Ha Hlai) via 23 autosomal STRs included in the Huaxia() Platinum System. Mol Genet Genomic Med 8(10):e1462

    Article  CAS  Google Scholar 

  21. Johnson KL et al (2004) Interlaboratory comparison of fetal male DNA detection from common maternal plasma samples by real-time PCR. Clin Chem 50(3):516–521

    Article  CAS  Google Scholar 

  22. Hansen MH, Clausen FB, Dziegiel MH (2012) Increased Y-chromosome detection by SRY duplexing. Fetal Diagn Ther 31(3):185–190

    Article  Google Scholar 

  23. Grosjean F, Favre M, Castella V (2022) Comparison between MACSprep forensic sperm microbead kit and Erase Sperm Isolation kit for the enrichment of sperm fractions recovered from sexual assault samples. Int J Legal Med. https://doi.org/10.1007/s00414-022-02861-7

    Article  Google Scholar 

  24. Wu D et al (2009) Validation of Differential Extraction Kit in forensic sexual assault cases. Fa Yi Xue Za Zhi 25(6):440–442

    CAS  Google Scholar 

  25. Iwanaga A et al (2008) Ablation of the scaffold protein JLP causes reduced fertility in male mice. Transgenic Res 17(6):1045–1058

    Article  CAS  Google Scholar 

  26. Ribas-Maynou J et al (2015) Nuclear degraded sperm subpopulation is affected by poor chromatin compaction and nuclease activity. Andrologia 47(3):286–294

    Article  CAS  Google Scholar 

  27. Chen J, Feng W, Zhan F (2019) Separation and forensic identification of sperm from cell mixtures using anti-hLCN6 monoclonal antibody coupled magnetic beads. Sheng Wu Gong Cheng Xue Bao 35(1):150–158

    CAS  Google Scholar 

  28. Huang G et al (2021) Removal of 1,2-benzanthracene via the intercalation of 1,2-benzanthracene with DNA and magnetic bead-based separation. Nucleosides Nucleotides Nucleic Acids 40(2):137–156

    CAS  Google Scholar 

  29. Hansenova Manaskova S et al (2016) Comparison of non-magnetic and magnetic beads in bead-based assays. J Immunol Methods 436:29–33

    Article  Google Scholar 

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Funding

This study was supported by Open Project of Shanghai Key Laboratory of Forensic Medicine and Judicial Expertise by the Ministry of Justice Key Laboratory (No. KF202110) and Basic Scientific Research Project of the Ministry of Public Security (No. 3242020014).

This study was approved by the Ethics Committee of China Medical University. All the participants were included after providing the written informed consent.

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Author notes

  1. Xiao-na Li and Feng-ling Xu are co-first authors.

Authors and Affiliations

  1. School of Forensic Medicine, China Medical University, No. 77 Puhe Road, Shenbei New District, Shenyang, 110122, China

    Xiao-na Li, Mao-ling Sun, Xiu-mei Zhu, Peng Lv, Zhe Du & Jun Yao

  2. Key Laboratory of Forensic Bio-Evidence Sciences, Liaoning Province, China

    Xiao-na Li, Mao-ling Sun, Xiu-mei Zhu, Peng Lv, Zhe Du & Jun Yao

  3. China Medical University Center of Forensic Investigation, Shenyang, China

    Xiao-na Li, Mao-ling Sun, Xiu-mei Zhu, Peng Lv, Zhe Du & Jun Yao

  4. Department of Forensic Medicine, China Criminal Police University, No. 38, Tawan Street, Huanggu District, Shenyang, 110854, Liaoning, China

    Feng-ling Xu & Ji-long Zheng

  5. School of Basic Medicine, China Medical University, Shenyang, 110122, China

    Xiu-peng Zhang

  6. Shanghai Key Lab of Forensic Medicine, Key Lab of Forensic Science, Ministry of Justice, China (Academy of Forensic Science), Shanghai, 200061, China

    Jun Yao

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  1. Xiao-na Li
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  2. Feng-ling Xu
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Correspondence to Xiu-peng Zhang or Jun Yao.

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Li, Xn., Xu, Fl., Zheng, Jl. et al. Magnetic bead-based separation of sperm cells from semen-vaginal fluid mixed stains using an anti-ACRBP antibody. Int J Legal Med 137, 511–518 (2023). https://doi.org/10.1007/s00414-022-02917-8

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  • DOI: https://doi.org/10.1007/s00414-022-02917-8

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