Hormonal and biochemical disturbances play an important role in development of infertility cases. Therefore, this study was conducted to evaluate the prolactin level and to explore the association between prolactin and thyroid stimulating hormone (TSH) and other hormones in the diagnosed infertile women in Ma'an governorate. The study was performed on 151 women (age group 20-40 years) who visited the Gynecology and Obstetrics Department at Ma'an governmental hospital. The study sample was divided into three groups: fertile group (n= 27), primary infertile group (n= 82) and secondary infertile group (n= 42). The results showed high incidence of prolactin abnormalities in both primary and secondary infertile women (90% and 92%, respectively). Furthermore, TSH abnormalities were found in 90% of primary infertile and % 93 of secondary infertile women. In addition, the results revealed the positively significant correlation between TSH and the fertility. Positive correlations were found between the levels of prolactin and TSH in primary and secondary infertility. Also, there were positive correlations between the levels of prolactin and other hormones, including luteinizing hormone, follicle-stimulating hormone, testosterone, estradiol and progesterone. In conclusion, the present study revealed a significant correlation between prolactin and TSH and other hormones in infertile women. Therefore, assessment of serum TSH and prolactin levels are mandatory in the work up of all infertile women, especially those presenting with menstrual irregularities.