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Updated: Sep 12, 2025
Flow Cytometric Analysis of Biomarkers for Detecting Human Sperm Functional Defects
Published on: April 21, 2022
[Different diagnostic methods for necrospermia and correlation between the severity of sperm necrosis and sperm DNA
Lang Fan1, Guo-Qiong Zhang2, Yu-Qiu Tan3
1Reproductive Center，Maternal and Child Health Hospital of Hunan Province, Changsha, Hunan 410008, China.
Objective:
To assess the value eosin-nigrosine (E-N) staining and hypo-osmotic swelling test (HOST) in the diagnosis of necrospermia and investigate the correlation of the degree of sperm necrosis with sperm DNA fragmentation index (DFI) and high DNA stainability (HDS).
Methods:
Using computer-assisted sperm analysis (CASA), we examined the sperm motility of 7 333 males seeking medical care in Maternal and Child Health Hospital of Hunan Province from May 2023 to July 2024, detected the sperm viability of those with asthenozoospermia by E-N staining and HOST, and measured the sperm DFI and HDS of those with necrospermia by sperm chromatin structure assay (SCSA). Based on sperm progressive motility (PR%), we divided the asthenozoospermia patients into three groups (PR% 20%－<30%, PR% 10%－<20% and PR% <10%), assessed the value of E-N staining and HOST in the diagnosis of necrospermia, and analyzed the correlation of the severity of sperm necrosis with sperm DFI and HDS.
Results:
Asthenozoospermia was diagnosed in 1 374 (18.74%) of the 7 333 males. The incidence rates of necrospermia revealed by E-N staining in the mild, moderate and severe asthenozoospermia groups were 0.55% (5/913), 3.80% (12/316) and 35.86% (52/145), respectively, with a total incidence of 0.94% (69/7333), while those detected by HOST were 0.99% (9/913), 6.96% (22/316) and 46.21%(67/145), respectively, with a total incidence of 1.34% (98/7333). There were no statistically significant differences in the results of diagnosis between the two methods (χ2 = 0.97, P > 0.05). Both E-N staining and HOST showed that sperm DFI in the necrospermia males was negatively correlated with sperm viability (r =－0.366, r=－0.333, P < 0.05), and so was sperm HDS, though with no statistically significant difference (P > 0.05).
Conclusion:
For males with PR％ <30%, sperm viability test should be conducted, with E-N staining as the choice of priority for the diagnosis of necrospermia. Sperm necrosis and sperm nuclear chromatin damage may be interactive risk factors.
