Development of a Noble On-Site Dual Screening System for Male Fertility of Sperm Count and Hyaluronidase Activity.

Purpose: To develop a noble on-site dual screening system for sperm concentration and hyaluronidase (HAdase) activity of male fertility based on lateral-flow assay methods, and to evaluate the effectiveness of this dual screening system.

Materials and methods: A chromogenic reaction for sperm count was established using a sperm-specific antibody against sperm antigen-10 (SP-10). Substrate-enzyme reaction was developed in which the activity of sperm HAdase on hyaluronic acid (HA) substrates could be measured through a color reaction. The colored HA substrates were degraded and the color faded by the sperm HAdase. To evaluate the validity of the developed methods, the results were compared using 49 semen samples (normozoospermia, 20; oligozoospermia, 9; azoospermia, 20) whose sperm counts were confirmed by the conventional method.

Results: Using color reaction by SP-10, no signal was observed in azoospermia and low count of sperm (<16×10⁶/mL), and the color signal increased as the count of sperm increased in normal and high count of sperm (≥16×10⁶/mL). The assay system for sperm HAdase activity was successfully developed. In the HAdase assay, the stronger the enzyme activity, the lower the color signal. Through statistical analysis, it was confirmed that there was a statistically significant correlation between sperm count and HAdase activity.

Conclusions: It was successfully developed that a novel on-site dual screening system for sperm count and HAdase activity, simultaneously. We are expecting that this novel dual assay system will provide the accessible and useful information about male fertility at on-site or home.