DNA Identification of Sperm Cells Collected and Sorted by Flow Cytometry

In cases of rape, obtaining enough biologic material for DNA identification of the attacker is often difficult because the methods for distinguishing and separating sperm cells from vaginal cells are not sufficiently efficacious. This article describes a new, innovative method for spermatic DNA extraction from the vaginal washing fluid by means of flow cytometry. The high specificity and sensitivity of the flow-cytometric sorting method provides enough sperm cells for DNA typing. The ease of execution of this method, involving vaginal washing with physiologic solution and flow-cytometric reading of the fresh sample, substantially increases its cost-benefit ratio.

[1]  L. Viola,et al.  Distribution of HumvWA31, HumFESFPS, HumTH01, HumTPOX, HumCD4, HumCSF1PO alleles in a Southern Italian population sample. , 2000, Journal of forensic sciences.

[2]  R. Sparkes,et al.  Validation of the AMPFlSTR SGM plus system for use in forensic casework. , 2000, Forensic science international.

[3]  I. Thorneycroft,et al.  Modified guanidinium thiocyanate method for human sperm DNA isolation. , 1997, Molecular human reproduction.

[4]  H. Sato,et al.  The modified method of two-step differential extraction of sperm and vaginal epithelial cell DNA from vaginal fluid mixed with semen. , 1995, Forensic science international.

[5]  B Budowle,et al.  Italian population data on two new short tandem repeat loci: D2S1338 and Penta E. , 1999, Forensic science international.

[6]  M. Klintschar,et al.  Separation of sperm and vaginal cells with flow cytometry for DNA typing after sexual assault. , 1999, Obstetrics and gynecology.

[7]  G. Makkar,et al.  Comparison of two colloidal silica-based sperm separation media with a non-silica-based medium. , 1999, Fertility and sterility.

[8]  B Budowle,et al.  Concordance study on population database samples using the PowerPlex 16 kit and AmpFlSTR Profiler Plus kit and AmpFlSTR COfiler kit. , 2001, Journal of forensic sciences.

[9]  T. Takatori,et al.  Isolation of spermatozoa in vaginal contents by centrifugation in a colloidal silica gradient. , 1980, Forensic science international.

[10]  A. Bongso,et al.  Comparative evaluation of two density gradient preparations for sperm separation for medically assisted conception. , 1999, Human reproduction.

[11]  B Budowle,et al.  CODIS STR loci data from 41 sample populations. , 2001, Journal of forensic sciences.

[12]  G. Dietl,et al.  DYS STR analysis with epithelial cells in a rape case. , 2001, Forensic science international.

[13]  Q. Schiermeier.  Y-chromosome analysis urged for sex crimes , 2001, Nature.

[14]  P. Wiegand,et al.  DNA extraction from mixtures of body fluid using mild preferential lysis , 2005, International Journal of Legal Medicine.

[15]  A. Amorim,et al.  Molecular characteristics of four human Y-specific microsatellites (DYS434, DYD437, DYS438, DYS439) for poulation and forensic studies. , 2001, Annals of human genetics.

[16]  B Budowle,et al.  STR primer concordance study. , 2001, Forensic science international.

[17]  B Budowle,et al.  Validation of short tandem repeats (STRs) for forensic usage: performance testing of fluorescent multiplex STR systems and analysis of authentic and simulated forensic samples. , 2001, Journal of forensic sciences.

[18]  R. Menkveld,et al.  Evaluation of three substitutes for Percoll in sperm isolation by density gradient centrifugation. , 1998, Human reproduction.

[19]  Regional Italian allele frequencies at nine short tandem repeat loci. , 2001, Forensic science international.

[20]  P. Walsh,et al.  Practical applications of genotypic surveys for forensic STR testing. , 2000, Forensic science international.

[21]  E. Fugger.  Clinical experience with flow cytometric separation of human X- and Y-chromosome bearing sperm. , 1999, Theriogenology.

[22]  P. Walsh,et al.  Sequence variation in humans and other primates at six short tandem repeat loci used in forensic identity testing. , 2001, Forensic science international.

[23]  R. Long,et al.  Separation of sperm through a 12-layer percoll column decreases the percentage of sperm staining with quinacrine. , 2000, Archives of andrology.

[24]  D. Strunk,et al.  Separation of sperm and vaginal cells based on ploidy, MHC class I-, CD45-, and cytokeratin expression for enhancement of DNA typing after sexual assault. , 1999, Cytometry.