title: Fluorescence- and magnetic-activated cell sorting strategies to separate spermatozoa involving plural contributors from biological mixtures for human identification.
abstract: No effective method has been developed to distinguish sperm cells originating from different men in multi-suspect sexual assault cases. Here we combined MACS and FACS to isolate single donor sperm cells from forensic mixture samples including female vaginal epithelial cells and sperm cells from multiple contributors. Sperms from vaginal swab were isolated by MACS using FITC-conjugated A kinase anchor protein 3 (AKAP3) antibody; target individual sperm cells involving two or three donors were separated by FACS using FITC-labeled blood group A/B antigen antibody. This procedure was further tested in two mock multi-suspect sexual assault samples and one practical casework sample. Our results showed that complete single donor STR profiles could be successfully obtained from sperm/epithelial cell and sperm mixtures from two contributors. For unbalanced sperm/epithelial cells and sperm cells mixtures, sensitivity results revealed that target cells could be detected at as low as 1:32 and 1:8 mixed ratios, respectively. Although highly relies on cell number and blood types or secretor status of the individuals, this procedure would still be useful tools for forensic DNA analysis of multi-suspect sexual assault cases by the combined use of FACS and MACS based on sperm-specific AKAP3 antigen and human blood type antigen.
authors.affiliation: Shanghai Key Laboratory of Crime Science Evidence, Key Laboratory of Forensic Evidence and Science Technology, Ministry of Public Security, Institute of Forensic Science, Shanghai Public Security Bureau, Shanghai 200083, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Department of Forensic Medicine, Shanghai Medical College, Fudan University, 138 Yixueyuan Road, Shanghai 200032, China.
authors.forename: Jianhui
authors.initials: J
authors.lastname: Xie
authors.affiliation: Shanghai Key Laboratory of Crime Science Evidence, Key Laboratory of Forensic Evidence and Science Technology, Ministry of Public Security, Institute of Forensic Science, Shanghai Public Security Bureau, Shanghai 200083, China.
authors.forename: Ronghua
authors.initials: R
authors.lastname: Chen
authors.affiliation: Shanghai Key Laboratory of Crime Science Evidence, Key Laboratory of Forensic Evidence and Science Technology, Ministry of Public Security, Institute of Forensic Science, Shanghai Public Security Bureau, Shanghai 200083, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Cao
authors.affiliation: Shanghai Key Laboratory of Crime Science Evidence, Key Laboratory of Forensic Evidence and Science Technology, Ministry of Public Security, Institute of Forensic Science, Shanghai Public Security Bureau, Shanghai 200083, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Ping
authors.affiliation: Shanghai Key Laboratory of Crime Science Evidence, Key Laboratory of Forensic Evidence and Science Technology, Ministry of Public Security, Institute of Forensic Science, Shanghai Public Security Bureau, Shanghai 200083, China.
authors.forename: Qingwen
authors.initials: Q
authors.lastname: Xu
authors.affiliation: Shanghai Key Laboratory of Crime Science Evidence, Key Laboratory of Forensic Evidence and Science Technology, Ministry of Public Security, Institute of Forensic Science, Shanghai Public Security Bureau, Shanghai 200083, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Hu
authors.affiliation: Shanghai Key Laboratory of Crime Science Evidence, Key Laboratory of Forensic Evidence and Science Technology, Ministry of Public Security, Institute of Forensic Science, Shanghai Public Security Bureau, Shanghai 200083, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Wu
authors.affiliation: Shanghai Key Laboratory of Crime Science Evidence, Key Laboratory of Forensic Evidence and Science Technology, Ministry of Public Security, Institute of Forensic Science, Shanghai Public Security Bureau, Shanghai 200083, China.
authors.forename: Lihua
authors.initials: L
authors.lastname: Gu
authors.affiliation: Shanghai Key Laboratory of Crime Science Evidence, Key Laboratory of Forensic Evidence and Science Technology, Ministry of Public Security, Institute of Forensic Science, Shanghai Public Security Bureau, Shanghai 200083, China.
authors.forename: Huaigu
authors.initials: H
authors.lastname: Zhou
authors.affiliation: Shanghai Key Laboratory of Crime Science Evidence, Key Laboratory of Forensic Evidence and Science Technology, Ministry of Public Security, Institute of Forensic Science, Shanghai Public Security Bureau, Shanghai 200083, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Chen
authors.affiliation: Department of Forensic Medicine, Shanghai Medical College, Fudan University, 138 Yixueyuan Road, Shanghai 200032, China.
authors.forename: Ziqin
authors.initials: Z
authors.lastname: Zhao
authors.affiliation: School of Life Sciences, Fudan University, 2005 Songhu Road, Shanghai 200438, China.
authors.forename: Jiang
authors.initials: J
authors.lastname: Zhong
authors.affiliation: School of Life Sciences, Fudan University, 2005 Songhu Road, Shanghai 200438, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Li
mesh_terms: D000017:ABO Blood-Group System; D000328:Adult; D056667:Biometric Identification / Q000379:methods*; D004847:Epithelial Cells / Q000166:cytology; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D005554:Forensic Medicine / Q000379:methods*; D006801:Humans; D008280:Magnetics; D008297:Male; D013094:Spermatozoa / Q000166:cytology*
chemical_list: D000017:ABO Blood-Group System