Developmental Validation of the SPERM HY-LITERTM Kit for the Identification of Human Spermatozoa in Forensic Samples


  • Presented in part at the 19th International Symposium on Human Identification, October 13–16, 2008, in Hollywood, CA.

  • The SPERM HY-LITER™ kit was developed and is manufactured by Independent Forensics of Illinois (IFI), a company engaged in the delivery of products and services to the forensic community. The Forensic Biotechnology Institute of California (FBIC) is a nonprofit ancillary unit of the College of Science and Mathematics at the California State University, Fresno. IRB permission was obtained for all samples reported in this study and taken by personnel at the California State University, Fresno, for studies conducted by University personnel at the University. Samples collected by personnel at Independent Forensics of Illinois (IFI) and tested at IFI by IFI personnel were not subject to IRB regulation.

Additional information and reprint requests:
Karl Reich, Ph.D.
Independent Forensics of Illinois
500 Waters Edge
Lombard, IL 60148


Abstract:  With sexual assault evidence, the visualization of spermatozoa confirms that ejaculation has occurred. However, microscopic examination of spermatozoa is a laborious process and can sometimes result in sperm cells being overlooked. Here, we present the developmental validation of the SPERM HY-LITER™ kit, which contains a human sperm–specific mouse monoclonal antibody coupled to a fluorescent Alexa 488 dye. The kit was tested using samples of human semen, saliva, blood, and urine, various animal semen extracts, sexual lubricants, and a commercially available spermicidal film. Postcoital vaginal swabs, degraded semen samples, and samples prepared with sample fixation techniques that deviated from the kit-provided protocol were also tested. In each case, the SPERM HY-LITER™ kit was demonstrated to bind only to human sperm cell heads. Limitations to this fluorescent staining procedure include nonspecific staining and increased background fluorescence with extreme heat fixation in some samples.