HY-LITER KITS
Introduction
SPERM HY‐LITER uses a fluorescently tagged anti-human sperm head monoclonal antibody to detect the presence of human sperm. Many identification methods for semen are directed towards protein markers in seminal fluid rather than to human sperm cells. Further, cell stains commonly relied upon to identify sperm provide no species information. SPERM HY‐LITER provides specific identification of the human origin of the sample and confirms that male‐origin cells are present.
Principle of the Test
SPERM HY‐LITER uses an Alexa 488 derivatized mouse monoclonal antibody to human sperm heads to specifically identify human sperm from sexual assault evidence by fluorescence microscopy. The method requires a fluorescence microscope: processed slides must be visualized on a fluorescence microscope fitted with the correct excitation and emission filters and light source.
In addition to a human sperm specific reagent, SPERM-HYLITER incorporates a second fluorescent dye that stains all
nuclei present in the sample (4′,6‐diamidino‐2‐
phenylindole, DAPI). Visualization of fluorescent nuclei is
not required for sperm detection, but is recommended for both manual and automated sperm searches.
Visualization of Human Sperm Heads
Cell nuclei, including epithelial and sperm, can be visualized using DAPI‐compatible filters. Human sperm heads can be visualized using fluorescein or Alexa 488 compatible filters. Slides may be scanned at a final magnification of 100x,
200x, or 400x at the operator’s discretion.
Specificity
SPERM HY‐LITER is specific for human sperm heads.
No cross‐reactivity with epithelial nuclei, blood cells or animal semen from horse, bull, sheep, goat, pig, dog,
cat, mouse and chimpanzee has been observed.
To date, semen from 3 nonhuman primates has been
tested: common chimp, Rhesus macaque and
cynomolgus macaque. SPERM HY‐LITER does NOT
detect sperm heads from these species.