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Updated: Aug 5, 2026
Flow Cytometric Analysis of Biomarkers for Detecting Human Sperm Functional Defects
Published on: April 21, 2022
The Association of DNA Fragmentation Index, Protamine Deficiency and Basic Semen Parameters: A Cross-sectional
Dimitrios Agkas1, Ioannis Tentes2, Ioannis Vakalopoulos3
1Laboratory of Physiology, Department of Medicine, School of Health Sciences, Democritus University of Thrace, Alexandroupolis, Greece.
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:
Elevated DFI and CMA3 were significantly and inversely correlated associated with total motility, progressive motility, vitality and normal morphology (P < 0.01), while DFI correlated positively with CMA3 positivity (ρ = 0.54, P < 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.
