The aim of the study was to determine if serial maternal urine polymerase chain reaction (PCR) tests can detect primary CMV infection during pregnancy. This was a prospective study conducted from 1 January 1999 to 31 December 1999 in an antenatal clinic setting of a teaching hospital. The study group included women who were CMV IgG negative and aged <30 years or had a pre-school child. They were invited to self-collect urine samples monthly and send them to the laboratory by post. Cord bloods were tested for CMV IgG to detect seroconversion. An anxiety questionnaire was sent to all study participants. At first attendance, 1549 (42%) women were CMV IgG negative. Of the 696 eligible women, 609 (88%) participated in the urine PCR study. PCR was performed on 2263 urine samples (median of 4/pregnancy). Primary CMV infection was identified in one woman by urine PCR at 36 weeks (baby CMV negative). Cord blood samples were available from 152/609 infants (25%). Seroconversion was noted in only one woman. Replies to the questionnaire were received from 264/609 women (43%): 214 (81%) had little or no anxiety, and 220 (83%) felt reassured by their study participation. Serial urine PCR is a feasible method of detecting primary maternal CMV infection during pregnancy which has potential for evaluation in further studies.