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DNA profiling of single sperm cells after whole genome amplification

  • Intimate swabs taken for examination in sexual assault cases typically yield mixtures of sperm and epithelial cell types. While powerful, differential extraction protocols to overcome such cell type mixtures by separate lysis of epithelial cells and spermatozoa can still prove ineffective, in particular if only few sperm cells are present or if swabs contain sperm from more than one individual leading to complex low level DNA mixtures. A means to avoid such mixtures consists in the analysis of single micromanipulated sperm cells. However, the quantity of DNA from single sperm cells is not sufficient for conventional STR analysis. Here, we describe a simple method for micromanipulating individual sperm cells from intimate swabs and show that whole genome amplification can generate sufficient amounts of DNA from single cells for subsequent DNA profiling. We recovered over 80% of alleles of haploid autosomal STR profiles from the majority of individual sperm cells. Furthermore, we demonstrate that in mixtures of sperm from two contributors, Y-STR and X-STR profiles of individual sperm cells can be used to sort the haploid autosomal profiles to develop the diploid consensus STR profiles of the individual donors. Finally, by analysing single sperm cells from mock sexual assault swabs with one or two sperm donors, we showed that our protocols enabled the identification of the unknown male contributors.

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Metadaten
Document Type:Article
Language:English
Author:Glenn M. G. Theunissen, Andrew Gibb, Paul Kong Thoo Lin, Burkhard Rolf, Sophia Forat, Richard Jäger
Parent Title (English):Forensic Science International: Reports
Volume:4
Article Number:100240
Pagenumber:10
ISSN:2665-9107
URN:urn:nbn:de:hbz:1044-opus-59674
DOI:https://doi.org/10.1016/j.fsir.2021.100240
Publisher:Elsevier
Publishing Institution:Hochschule Bonn-Rhein-Sieg
Date of first publication:2021/10/16
Copyright:© 2021 The Author(s). Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license.
Funding Information:This work was funded by the German Federal Ministry of Education and Research (project no. 03FH019PX4). Further support was provided by a PhD scholarship to GMGT from the Institute of Safety and Security Research (ISF) of Bonn-Rhein-Sieg University of Applied Sciences.
Keyword:DNA typing; Forensic genetics; Short tandem repeat (STR); Whole genome amplification; X-STR; Y-STR; micromanipulation; sexual assault; sperm cell
Departments, institutes and facilities:Fachbereich Angewandte Naturwissenschaften
Institut für Sicherheitsforschung (ISF)
Institut für funktionale Gen-Analytik (IFGA)
Projects:ForWArd - FHprofUnt 2014: Forensische Analyse singulärer Genome mittels WGA (DE/BMBF/03FH019PX4;13FH019PX4)
Dewey Decimal Classification (DDC):5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 576 Genetik und Evolution
Entry in this database:2021/10/21
Licence (German):License LogoCreative Commons - CC BY-NC-ND - Namensnennung - Nicht kommerziell - Keine Bearbeitungen 4.0 International