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Updated: Aug 5, 2026
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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.
Journal of Human Reproductive Sciences
|August 4, 2026
Summary
Sperm DNA Fragmentation Index (DFI) and protamine deficiency (CMA3) correlate with reduced semen quality. These molecular markers offer valuable insights into male infertility beyond conventional semen analysis.
Area of Science:
- Andrology and Reproductive Medicine
- Molecular Biology and Genetics
- Clinical Diagnostics
Background:
- Conventional semen analysis may not detect sperm nuclear integrity defects.
- Molecular markers like DNA Fragmentation Index (DFI) and protamine deficiency offer complementary insights into sperm chromatin quality.
Purpose of the Study:
- To investigate associations between conventional semen parameters and DFI.
- To evaluate the relationship between semen parameters and protamine deficiency using chromomycin A3 (CMA3) staining.
Main Methods:
- Cross-sectional analytical study in a certified andrology laboratory.
- Included 150 men undergoing fertility evaluation (2022-2024).
- Assessed DFI (Halosperm) and CMA3 staining alongside WHO 2021 semen analysis.
Main Results:
- Elevated DFI and CMA3 showed significant inverse correlations with total motility, progressive motility, vitality, and normal morphology (P < 0.01).
- DFI positively correlated with CMA3 positivity (ρ = 0.54, P < 0.001).
- Men with DFI >30% or CMA3 >40% exhibited significantly reduced semen quality.
Conclusions:
- DFI and CMA3 are strongly associated with key semen parameters.
- These markers provide complementary information on sperm nuclear integrity.
- DFI and CMA3 hold potential clinical relevance in diagnosing male infertility.
