Optimization of semen antioxidant capacity, sperm DNA damage and sperm mitochondrial mutation analysis methods

Infertility, defined by the World Health Organization (WHO) as an unprotected sex after one year of unprotected sexual intercourse, affects 10-15% of couples in developed and developing countries. However, since semen analysis used in the diagnosis of routine infertility provides diagnosis at a very low rate, other additional techniques, new approaches and more comprehensive combined testing are needed in diagnosing cases of infertility. The present study also aimed to be used for diagnostic purposes in non- varicocele infertile men; sperm biomarkers panel including semen analysis, semen- antioxidant capacity, nDNA packaging and fragmentation, and mtDNA damage. The analysis methods of the parameters to be tested in the panel were optimized and evaluated in 5 normal and 2 infertile male patients. The parameters in the panel are; semen analysis based on WHO criteria, mRNA damage was assessed using the SORP analysis, nDNA packaging and fragmentation TUNEL and Aniline blue tests using the SORP analysis, mRNA DNA damage was assessed using the PZR "common deletion" gene region and the ATPase6, ND3, amplification. Study results; only 6 infertile patients showed low sperm count and high sORP. In the presented thesis study, optimization of analysis methods to be used for a panel study consisting of four parameters with the combined test approach has been provided for the diagnosis of male infertility. Completion of the optimization studies for the techniques to be applied has opened and facilitated the evaluation of future planned infertile male patients. In addition, the evaluation of these parameters as a whole will ensure that the failed pregnancy process and assisted reproductive techniques are not experienced. Keywords: Male infertility, sperm, mtDNA, deletion, oxidative stress, motility, ROS, sperm DNA