Background: central line-associated blood stream infection (CLABSI) is responsible for almost 60% of the hospital-acquired infections. The rate of CLABSI in developing countries is higher than that in developed countries. Recently; gram negative bacilli are commonly isolated in CLABSI cases. Aim of the work: This was a prospective 6 month study to assess the occurrence of CLABSI at Pediatric Intensive Care Unit (PICU) of Ain-shams University Hospital. Patients and methods: present study was a prospective observational study conducted on 109 patients with a 152 central venous catheter (CVCs) inserted to them, who were admitted to the Pediatric Intensive Care Unit of Ain Shams University Hospital from July to December 2017. They were observed for development of central lineassociated blood stream infection (CLABSI) and also associated risk factors were studied. Results: There was 22 CLABSI cases with a CLABSI score 14.1 per 1000 central line days. In this study, the most common organism detected was Klebsiella that was isolated in 14 cases. This study shows multi-drug resistant pathogens as causative agent of CLABSI in 9 % of cultures. Conclusion: CLABSI is a common and serious complication to CVCs in PICU. Klebsiella, candida albicans are the common pathogens isolated in CLABSI.