Facing a 1.2% HIV-1 seroprevalence amongst its adult female population, Cambodia recently established national guidelines for the expansion of services to prevent mother-to-child transmission. Anticipating this expansion, Sihanoukville Hospital performed an anonymous HIV-1 cord-blood serosurvey of 600 consecutive deliveries from May 2001 through June 2002. Laboratory analysis utilizing ELISA and Western blot techniques yielded a 4.2% seroprevalence; a value resembling previous reports from other Sihanoukville antenatal clinics (3.9%) but exceeding the national ANC clinic average of 2.3%. Demographic information was confidentially collected and analysed for co-variation to HIV-1 status. One hundred percent of HIV positive mothers (P =0.013) self-reported the occupation of housewife, consistent with previous documentations suggesting the shift in infection burden from high-risk groups into the general female population. A substantial proportion received no prenatal care, suggesting that interventions addressing mother-to-child transmission will need to prepare for the management of pregnant women presenting at or near the time of delivery.