Enterobacter sepsis is commonly recognized as a hospital-acquired infection in childhood. In a five year prospective surveillance of neonatal sepsis at the Aga Khan University Hospital in Karachi, we identified Enterobacter sepsis in 28/292 (10%) cases, with an incidence of 0.7 per thousand births among inborn infants. There was no significant difference in predisposing factors and clinical features between Enterobacter and other infections. Approximately half (47%) of Enterobacter infections presented within 72 h of birth and the associated mortality was 21%. Increasing resistance to commonly used first- and second-line antibiotics over the last five years was noted. Enterobacter infections are emerging as significant pathogens among cases of neonatal sepsis in Karachi.