Objectives: To observe frequency of Staphylococcus epidermidis in suspected septicemic newborns. This is a cross-sectional descriptive study conducted in Shaheed Mohtrama Benazir Bhutto Medical College in department of pathology with collaboration of Dept. of pediatric Sind Government Lyari General tertiary care hospital of Karachi. from June 2012 to December 2012. Material & Methods Blood samples for culture were collected from 105 newborn admitted in Neonatal Intensive Care Unit with suspected septicemia. Specimens were inoculated in brain heart infusion broth in ratio of 1:5. Subcultures were performed at 24 hours, 48 hours &72 hours and on the 7th day. Isolates were identified by standard methods. Result: A total of 70, organisms isolated out of 105 samples. These include Klebsiella species Which were 18, Staphylococcus aureus were 10, Staphylococcus epidermidis were 13, Proteus mirabilis 7, Enterobacter species 5, Pseudomonas species 6, Escherichia coli 5, Acinetobacter species 3, Citrobacter species 2, and Staphylococcus saprophyticus 1. Conclusion: In this study Staphylococcus epidermidis tends to show a major causative pathogen with higher frequency now a day in neonates.