Background: Sepsis in neonates (NS) is a global issue that possesses a challenge for management by pediatrics health care providers. Objective: Study the clinical and bacteriological profile of 80 preterm neonates. This was a case controlled study conducted at Almadina Almunawara, Saudi Arabia on 80 preterm neonates. Patients and Methods: The included neonates were divided into 2 equal groups, the first groups included 40 preterm neonates directed to neonatal intensive care unit (NICU) with diagnosis of sepsis that proved was with clinical and laboratory investigations and the second group included 40 healthy preterm neonates. Results: The gestational age and Apgar score were lower in neonatal sepsis group than normal group with a significant difference. The respiration rate was significantly increased in neonatal sepsis group than normal group. A significantly higher count of neutrophil and IT were found in the neonatal sepsis group in comparison with normal group. However, platelets count was lower in NS group than normal group. The most prevalent culture isolate among neonatal sepsis group were gram positive bacteria as CONS (27.5%) and staph aureus (20%). The gram negative bacteria were klebsiella Pneumonia (20.8%) followed by E coli (15%) then Acenobacter (10%) and pseudomonas (10%). Conclusion: Preterm pregnancy and CS delivery is associated with higher risk for neonatal sepsis. Gram positive bacteria were the most prevalent type of isolates in our hospital including CONS and staph aureus. The gram negative bacteria were klebsiella Pneumonia followed by E coli then Acenobacter and pseudomonas (10%).