Infertility is a major public health problem worldwide and its prevalence is as high as 17, 5% depending on the population studied The research is aimed to study the possibility of using miR-20a, miR-135a, miR-34b, miR-449v, miR-449c as markers of male infertility and to evaluate the dependence of the effectiveness of ART programs on the level of expression of exosomal microRNAs. The experimental group included patients entering the program of ART (assisted reproductive technologies) with the diagnosis of idiopathic male infertility (n = 30), the control group included couples with female infertility of tubal origin (n = 19). The isolation of exosomal microRNA from ejaculate was performed using the exoRNeasy Midi regent kit. The miRCURY LNA miRNA SYBR® Green PCR System was used to evaluate the expression of the following exosomal microRNAs: miR-20a, miR-135a, miR-34b, miR-449c, miR-449c and control miR-16. The expression of exosomal miR-449c and miR-135a was significantly different in the experimental group (p = 0.03 and p = 0.04, respectively). There was also a tendency to decrease the expression for such microRNAs as: miR-20a, miR-34b and miR-449c. Moreover, the expression level of miR-34b, miR-449c, and miR-449c and miR-135a has a direct correlation with the effectiveness of ART programs (p < 0.05 for miR-135a). The strength of correlation relationship of the above relationships using Cheddock scale was moderate. MicroRNA molecules selected for the study not only demonstrated their potential ability to be used as a diagnostic marker of male infertility, but also showed the ability to reflect the efficiency of fertilization and embryo formation processes.