Epstein-Barr virus (EBV) infection of the human uterine cervix was detected in 5 out of 28 women by means of culture and cytohybridization analysis of cervical secretions. Cervical samples from 2 of 14 women contained epithelial cells with EBV DNA, and filtered cervical washings from 4 women contained infectious EBV. The discovery of EBV shedding in its cell-free infectious form from the uterine cervix raises the possibility of venereal transmission, neonatal infection, and EBV involvement in cervical pathology.