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Background. Kisspeptin system plays an important role in the neuroendocrine control of gonadotropin secretion, sexual differentiation of the brain, puberty, and fertility. Objective — the study was aimed at assessing blood level of Kisspeptin in healthy boys, depending on puberty stage, as well as in boys with pathological delay of puberty onset. Material and methods. The study included 43 somatically healthy boys. Group 1 consisted of 12 boys 14—17 years old with delayed puberty, group 2 included 16 boys 14—18 years old with normal puberty, Tanner stage IV — V, group 3 — 15 boys 6—10 years old, Tanner stage I. In group 1, median testosterone level (T) was 0.09 nmol/l, LH — 0.3 IU/l; the peak LH level in the triptorelin test was below 5 IU/l in 50% of this group. Serum level of kisspeptin was assessed by ELISA using a specific kit. Results. Median serum level of kisspeptin (34.8 pg/ml) was significantly (p>0.05) higher in in the group with delayed puberty than in the groups with age-appropriate sexual development. There were no differences between groups 2 and 3 in this value. Conclusion. Serum level of kisspeptin, which is low in the case of physiological sexual development (regardless of stage), significantly increases in boys with delayed puberty onset, which can be used as a diagnostic marker of this pathology. The results suggest attenuated effect of kisspeptin due to decrease in its biological activity or decrease in sensitivity of the kisspeptin receptors in the case of delayed puberty and hypogonadism. The possibility of treatment of male hypogonadism with exogenous kisspeptin cannot be ruled out.