The utility of sperm DNA damage assay using toluidine blue and aniline blue staining in routine semen analysis

Clin Exp Reprod Med. 2013 Mar;40(1):23-8. doi: 10.5653/cerm.2013.40.1.23. Epub 2013 Mar 31.

Abstract

Objective: The aim of the present study was to examine the relationship among male age, strict morphology, and sperm chromatin structure and condensation.

Methods: Sperm samples from a total of 100 men underwent semen analysis, and sperm chromatin structure and condensation were assessed with toluidine blue (TB) and aniline blue (AB) tests.

Results: Prevalence of strict morphology of less than 4%, and abnormal sperm chromatin structure and condensation did not show any statistically significant differences according to male age (p=0.605, p=0.235, and p=0.080). No significant correlation was demonstrated among age of male partners, strict morphology, and abnormal sperm chromatin structure using TB and AB tests. However, abnormal sperm chromatin condensation was positively associated with sperm chromatin structure (r=0.594, p=0.000) and showed negative correlation with strict morphology (r=-0.219, p=0.029).

Conclusion: The tests for sperm chromatin condensation showed a significant association with strict morphology. Further study is needed to elucidate the relationship between clinical outcome and sperm chromatin tests.

Keywords: Aniline blue; DNA damage; Human; Semen analysis; Toluidine blue.