HBA as a Potential Alternative to the TUNEL Assay for Assessment of DNA Integrity in Asthenozoospermic Spermatozoa

Abstract

Objective: This study was conducted to evaluate the appropriateness of the hyaluronan binding assay (HBA) for assessment of DNA integrity indirectly in asthenozoospermic spermatozoa, by studying its correlation with the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Methods: Thirty asthenozoospermic semen samples of infertile couples were recruited. Semen analysis was performed before being processed by HBA and the TUNEL assay. All the semen parameters were recorded and analyzed. Results: The mean age of the patients was 38.1 ± 6.4 years. The average total and progressive motility was 33.5 ± 11.0% and 21.9 ± 6.6%, respectively. The mean TUNEL-positive and hyaluronan-bound sperm were 17.8 ± 6.4% and 57.2 ± 14.5%, respectively. Pearson correlation coefficient showed a high negative correlation between HBA and the TUNEL assay (r = -0.73, p < 0.001). The intraclass correlation coefficients revealed a high level of agreement between two observers who evaluated the TUNEL-positive and hyaluronan-bound sperm [0.81 (95% CI: 0.65–0.91) and 0.92 (95% CI: 0.84–0.96)], respectively. Conclusion: The study results showed a high negative correlation between HBA and the TUNEL assay. This suggests that HBA may be used as a viable intact DNA spermatozoa identification method, as a potential alternative to the TUNEL assay, in asthenozoospermia sperm.

Keywords

• Spermatozoa
• asthenozoospermia
• hyaluronan binding assay
• TUNEL assay
• infertility and DNA fragmentation
• (Siriraj Med J 2018;70: 70-76)