To determine if cytomegalovirus (CMV) DNA could be detected in the cerebrospinal fluid (CSF) of infants with congenital infection with CMV, polymerase chain reaction (PCR) was done on CSF samples from 13 infants (10 with confirmed, 1 with possible, and 2 with asymptomatic congenital CMV infection) and on CSF samples from 100 control patients with alternative diagnoses. By use of two sets of primers that targeted different areas of the CMV genome, CMV DNA was amplified in the CSF of 6 of 10 symptomatic infants, 0 of 3 infants with asymptomatic CMV infection or possible congenital CMV disease, and 2 control patients (one primer set only). A positive CSF PCR result at birth correlated with a poor neurodevelopmental outcome (P = .048; two-tailed Fisher's exact test).