Several diagnostic assays for neonatal and infantile chlamydial infections, isolation with tissue culture, antigen detection by enzyme immunoassay (EIA) (IDEIA Chlamydia test), a nonisotopic DNA probe (Gen-Probe PACE 2 assay), serum IgM antibody detection by EIA (SeroELISA Chlamydia TRUE IgM), and polymerase chain reaction (PCR), were evaluated. Of 210 clinical specimens (170 nasopharyngeal and 40 conjunctival swabs) from 53 neonates and 102 infants with respiratory insufficiency and respiratory tract infections which were suspected to be associated chlamydial infection, chlamydial antigens were detected in 30 by IDEIA Chlamydia. Of these 30 samples, C. trachomatis was isolated from 27 specimens. Samples from 15 neonates and 6 infants were culture-positive and IDEIA Chlamydia-positive. Of 30 samples, 27 were tested with PCR and 8 with DNA probe. Twenty-three of 27 specimens were positive with PCR, while only one specimen was positive with DNA probe. EIA can be used for the diagnosis and screening of neonatal and infantile chlamydial infections.