Using Sperm chromatin Staining Techniques as a Predictive Diagnostic Tool for Male Infertility

Abstract

Background : It is a scientific fact today that routine seminal analysis is only a poor predictor of male reproductive potentiality , so that there is an increased need for finding out new functional tests in the assessment of male infertility . Objective : The aim of the present study was to investigate the predictive value of sperm chromatin integrity assays as a complementary tool to assist in diagnosis of infertility . Method : This study was carried out between January 2013 and November 2013 including one hundred (100) selected infertile men who attended to fertility Center in Al-Sadr Medical City . The study also included twenty (20) healthy fertile volunteers as a control . Aniline blue (AB) staining was used to differentiate between normal condensed chromatin (unstained) and abnormal decondensed chromatin (blue), while toluidine blue (TB) was used to differentiate between normal intact sperm DNA (blue) and abnormal damaged sperm DNA (purple), this is the first study in Iraq that employed (TB) to assess sperm DNA fragmentation . Results : The results showed that there was a high significant difference (p<0.01) in sperm chromatin decondensation and sperm DNA damage between infertile and fertile men . The study also revealed a significant difference (p<0.05) in sperm DNA damage between normozoospermic and fertile men, but no significant difference (p>0.05) was found in the sperm chromatin condensation . Both sperm chromatin condensation and sperm DNA damage showed a significant positive correlation with sperm morphology, and a significant negative correlation with sperm concentration, motility and progressive motility. Conclusions : The study concluded that sperm DNA fragmentation (by TB staining) has good predictive value in discriminating infertile and fertile men over seminal fluid analysis . Recommendations : TB and AB staining of sperm chromatin can be used in the course of infertility diagnosis . However, more investigations may be needed to confirm the discriminating power of sperm chromatin condensation assays (by AB staining) to be ready to use in the routine workup in the fertility clinics .