DNA profiling of single sperm cells after whole genome amplification

Forensic Science International: Reports

Andrew Gibb¹  Sophia Forat²  Paul Kong Thoo Lin³  Glenn M.G. Theunissen⁴  Richard Jäger⁵  Burkhard Rolf⁵ 

[1] Institute of Safety and Security Research, Hochschule Bonn-Rhein-Sieg, University of Applied Sciences, Grantham Allee 20, 53757 Sankt Augustin, Germany;SPA Forensic Services, Nelson Street, Aberdeen, Scotland AB24 5EQ, UK;School of Pharmacy and Life Sciences, Robert Gordon University, Aberdeen AB10 7GJ, Scotland, UK;Department of Natural Sciences, Bonn-Rhein-Sieg University of Applied Sciences, von-Liebig Str. 20, 53359 Rheinbach, Germany;School of Pharmacy and Life Sciences, Robert Gordon University, Aberdeen AB10 7GJ, Scotland, UK;

关键词: Short tandem repeat (STR);    Forensic genetics;    DNA typing;    Whole genome amplification;    Y-STR;    X-STR;   

DOI  :  

来源: DOAJ

【 摘 要 】

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 analyzing 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.

【 授权许可】

Unknown