Diagnosis of HIV infection among children born to HIV-positive mothers can be made in the first 12 months, but few studies have examined HIV status during the first weeks of life. In a prospective longitudinal study of 50 infants born to HIV-1 seropositive women, blood samples were obtained at birth and at 4-9 weeks and 5-9 months of age, and were tested for HIV-1 by the polymerase chain reaction (PCR), viral culture, and p24 antigen measurements. 16 were diagnosed as HIV-infected by the age of 4-9 weeks according to both PCR and culture; by contrast, infection could be detected in only 5 children at birth. No changes in HIV status were observed between 4-9 weeks and 5-9 months in the 44 children who could be retested. Perinatal HIV-1 infection can therefore be diagnosed in the first 2 months of life, either by PCR or viral culture. Our inability to detect HIV-1 infection at birth in almost 70% of babies subsequently found infected suggests an active replication of HIV during the first weeks of life. Our results might favour the hypothesis that transmission of HIV-1 takes place either at the end of pregnancy or at delivery.