With the increased survival of VLBW infants (<1500 grams), Candidemia has become an important cause of morbidity and mortality in the NICU. The epidemiology of Candidal disease in the NICU, however, has not been well-studied, especially with regards to the latest DNA typing methods. Using pulse field gel electrophoresis (PFGE) of Candidal chromosomal DNA, we studied Candidal blood isolates over a 53 month period in a single level III NICU. During that time, there were 16 cases of Candidal fungemia proven by blood culture and one case identified with a positive CSF culture. These neonates had similar risk factors for systemic Candidal disease as had previously been described in the literature, including prematurity, prolonged antibiotic use, TPN and intralipid administration, prolonged endotracheal intubation, and the presence of indwelling vascular catheters. Of these 17 cases, 11 of the blood isolates and the CSF isolate were available for DNA typing. Typing revealed 4 strains of C. parapsilosis (6 blood isolates and the CSF isolate), 3 strains of C. albicans (4 blood isolates total), and a single blood isolate of C. guillermondii. One strain of C. parapsilosis was found in 4 infants spanning over 3 years. Another strain of C. albicans was found in two infants 2 months apart. All other strains of Candida were dissimilar. We suggest, using the sensitive and reproducible technique of PFGE, that Candida species causing systemic disease in the NICU can be acquired not only from the maternal genital tract, but also from nosocomial endemic sources spanning long periods of time. We propose that these results will help lay the epidemiologic groundwork for further study into the source, and ultimate prevention, of fungal disease in the nursery.