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 Science International. Genetics
Lee DeanChristopher J Ehrhardt

Abstract

Analysis of biological mixtures is a significant problem for forensic laboratories, particularly when the mixture contains only one cell type. Contributions from multiple individuals to biologic evidence can complicate DNA profile interpretation and often lead to a reduction in the probative value of DNA evidence or worse, its total loss. To address this, we have utilized an analytical technique that exploits the intrinsic immunological variation among individuals to physically separate cells from different sources in a mixture prior to DNA profiling. Specifically, we applied a fluorescently labeled antibody probe to selectively bind to one contributor in a mixture through allele-specific interactions with human leukocyte antigen (HLA) proteins that are expressed on the surfaces of most nucleated cells. Once the contributor's cells were bound to the probe, they were isolated from the mixture using fluorescence activated cell sorting (FACS)-a high throughput technique for separating cell populations based on their optical properties-and then subjected to STR analysis. We tested this approach on two-person and four-person whole blood mixtures where one contributor possessed an HLA allele (A*02) that was not shared by other contri...Continue Reading

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Citations

May 19, 2017·Analytical and Bioanalytical Chemistry·M Katherine PhilpottChristopher J Ehrhardt
Jan 24, 2017·F1000Research·Ye Jin KwonChristopher J Ehrhardt
Nov 7, 2019·The American Journal of Forensic Medicine and Pathology·Papanu Suttipasit
Dec 29, 2020·Science & Justice : Journal of the Forensic Science Society·Kaitlin HuffmanJack Ballantyne
Jan 12, 2020·Science & Justice : Journal of the Forensic Science Society·Agnieszka KuffelNiamh Nic Daeid
Mar 11, 2020·Forensic Science International. Genetics·Julia BurrillNunzianda Frascione

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