Healthcare-associated infections (HAI) are associated with increased morbidity, mortality and associated financial burden. Nosocomial blood stream infections in NICU remain a serious hazard. This was a prospective study done from January to December 2016. 1064 neonates admitted to NICU during this period were enrolled in the study. Active surveillance in NICU and laboratory based surveillance in Department of Microbiology was carried out to find cases of nosocomial blood stream infections. Each case was line listed and followed with calculation of attack and incidence rates. Out of the total 1064 new admissions between January and December 2016, 30 cases of hospital acquired infections were identified based on risk factors, clinical and laboratory data. Out of these twenty cases were hospital acquired blood stream infections. Average attack rates of HAI and hospital acquired blood stream infection were 2.76% and 1.93% respectively. Incidence of HAI/1000 patient days was 5.57 with BSI rate of 12.74. / 1000 catheter days. The most common organisms isolated from positive blood cultures were Staphylococcus aureus (32.4%), Acinetobacter spp. (24%) and E. coli (11.3%). BSI in NICU remains a serious hazard and remains grossly underreported. Infection control policies and surveillance methodologies need to be strengthened to curb BSI.