Despite the researchers’ efforts, the cause and development of breast cancer is still incompletely understood. Currently, in some reports, human cytomegalovirus has been referred as a risk factor for breast cancer. This study aimed to determine relative frequency of cytomegalovirus in tissue samples of women with breast cancer in Sanandaj County. In this study, to determine the relative frequency of the human cytomegalovirus (CMV) 50 formalin-fixed tissues of breast cancer, which all were invasive ductal carcinoma, were studied using the nested-polymerase chain reaction. In 26 cases of breast cancer tissues (26/50), human cytomegalovirus was detected. Seventeen cases of breast cancer tissues were in a moderately differentiated stage, and nine cases had poor-differentiated stage tissues that were positive for viral DNA. At older ages (>45 years) the prevalence rate of human cytomegalovirus DNA was higher, but no significant association was seen (p=0.16). In general, due to the high prevalence of the DNA of human cytomegalovirus (58%), in this study it is assumed that human cytomegalovirus (HCMV) has a contributing role in breast cancer; although more study is required to clearly define its part in this type of cancer.