ABSTRACT Context: The study helps to make antibiotic policy in neonatal septicemia.ObjectiveThe present study was undertaken to investigate the high incidence of multidrug resistant β lactamases producing Gram-negative bacilli causing neonatal septicemia along with their antimicrobial sensitivity pattern Design & SettingThe eighteen months prospective study was conducted from June 2011 to Dec 2012. The Blood samples from the suspected neonatal septicemias were processed in the Department of Microbiology SGRDIMSR Amritsar. After identification and antibiotic susceptibility testing, beta-lactamases were detected as per CLSI guidelines.ResultsIn 235 blood cultures 37.87% (89/235) positivity was seen. Out of them, 55.05% were Gram positive and 43.82% were Gram negative isolates while Candida was 1.12%.Among S. aureus 62.0% were MRSA while 37.93% were MSSA. CONS were 40.8%. Klebsiella (25.64%) was predominant gram negative isolate followed by Enterobacter (20.51%), Acinetobacter (17.94%), Pseudomonas (15.38%), E coli (12.8%) and Citrobacter (7.69%).69.23% were lactamases producer gram negative isolates. Maximum 80.0% was in Klebsiella followed by 71.42% Acinetobacter, 66.67% in Citrobacter and Pseudomonas spp 62.5% Enterobacter, and 60.0% in E coli. ESBL producers were maximum 45.74% followed by co-producers of MBL + AmpC 18.5%,ESBL+AmpC 14.8%, ESBL+MBL11.12%,AmpC and MBL 7.4%.ConclusionsThe varying microbiological pattern of neonatal septicaemia warrants the need for periodic review of neonatal sepsis as the knowledge of the pathogens and their antibiotic susceptibility would be a useful guide in the antibiotic therapy This will also facilitate infection control interventions like hand hygiene, patient isolation and contact precautions.