Blood drawn from neonatal patients is available in only limited amounts for blood cultures. The BACTEC Pediatrics (PEDS) Plus aerobic nonradiometric blood culture bottle was designed to aid the diagnosis of pediatric bacteremia. To assess the value of PEDS Plus blood culture medium for neonatal patients, we prospectively compared the PEDS Plus blood culture medium with the standard BACTEC aerobic medium (NR6A). From January to December 1999, 192 pairs of PEDS Plus and NR6A blood culture bottles were collected by the pediatric microbiology laboratory of Taipei Veterans General Hospital. Seventeen (8.85%) isolates were considered to be clinically significant microorganisms. Isolation rate of the PEDS Plus culture bottles was significantly higher than that of the NR6A culture bottles (p < 0.001). Six isolates were detected at least 1 day earlier by using the PEDS Plus culture bottles (p < 0.001). Among those patients who were under antibiotic therapy, the PEDS Plus culture bottle showed more significant growth than the NR6A culture bottle (p < 0.001). After weighing their benefits, we suggest substituting PEDS Plus bottles for NR6A bottles when culturing blood from neonatal patients or from those who are receiving antibiotic therapy.