The Association of DNA Fragmentation Index, Protamine Deficiency and Basic Semen Parameters: A Cross-sectional Analytical Study

Background: Conventional semen analysis may overlook defects in sperm nuclear integrity, whereas molecular markers such as DNA Fragmentation Index (DFI) and protamine deficiency provide complementary information on sperm chromatin quality. Aim: The aim of this study was to investigate the associations between conventional semen parameters and two clinically relevant markers of sperm chromatin integrity: DFI and protamine deficiency assessed by chromomycin A3 (CMA3) staining. Settings and Design: This was a cross-sectional analytical study conducted in a certified private andrology laboratory operating under standardised quality control protocols. Materials and Methods: A total of 150 men undergoing fertility evaluation between 2022 and 2024 were included. Semen analysis was performed according to the World Health Organization 2021 guidelines;[1] DFI was assessed using the Halosperm sperm chromatin dispersion test, and chromatin maturity was evaluated by CMA3 staining. Statistical Analysis Used: Non-parametric statistical methods were applied to evaluate correlations between molecular markers (DFI and CMA3) and semen parameters. Results: < 0.001). Men with DFI >30% or CMA3 >40% showed significantly reduced semen quality parameters. Conclusions: DFI and CMA3 are strongly associated with key semen parameters and provide complementary information on sperm nuclear integrity with potential clinical relevance in male infertility.