UTILITY OF DIFFERENT SPERM DNA FRAGMENTATION TESTS FOR THE ASSESSMENT OF MALE FACTOR AND REPRODUCTIVE OUTCOMES IN AUTOLOGOUS CYCLES

To study different techniques for measuring sperm DNA fragmentation and its correlation with semen quality and reproductive outcomes in assisted reproduction This prospective observational study, conducted from September 2022 to March 2023, enrolled 94 males seeking assisted reproductive treatment (IVF/ICSI) with their partners. Males with infectious and/or chronic diseases, or under drug treatment were excluded. Fresh semen samples were collected by masturbation, analyzed according to WHO criteria and separated in two aliquots. One of them was used for sperm DNA fragmentation analysis using three different techniques: TUNEL assay, sperm chromatin dispersion (SCD) test and double straining breaks (DSBs) detection by using phosphorylated H2AX antibody. The other aliquot was used for autologous IVF/ICSI cycles. Linear regression and Pearson correlation coefficients were used to analyze correlations between the different variables Following WHO criteria, 11.7% of the enrolled patients were oligozoospermic, 22.34% were asthenozoospermic, and 6.38% were oligoasthenozoospermic. The mean sperm DNA fragmentation index was 10.86% for TUNEL assay, 10.94% for DSBs, and 23.98% for SCD assay. Male age was significantly positive correlated with SCD score (p=0.0253). In addition, we observed relation between the DNA fragmentation index, measured by TUNEL and SCD essays, and seminal parameters. Negative correlations were found between DNA fragmentation and progressive motility (p<0.0001) for both TUNEL and SCD, and with total progressive sperm count (p=0.0262) for TUNEL assay, while no significant correlation was found between seminal parameters and DSBs. No relation was found between fertilization rate and good quality embryo rate with DNA fragmentation measured by any technique. Lastly, when reproductive outcome was analyzed, a significant negative correlation was found between sperm DNA fragmentation measured by DSBs and clinical pregnancy rate (p=0.0485), but not by TUNEL (p=0.7569) or by SCD (p=0.9307) Elevated sperm DNA fragmentation index determined by TUNEL and SCD assays reflects abnormalities in seminal quality. DSBs score did not reveal abnormalities in seminal parameters but it is associated with lower pregnancy rate