Abstract: Limited data exist on economic burden of rotavirus gastroenteritis in Libya. Aim of this study is to perform cost analysis of rotavirus disease in Libya. A prospective cross sectional study was conducted among rotavirus children aged below five years from August 2012 to April 2013 at three public hospitals in Libya. The cost of rotavirus treatment is calculated the cost of rotavirus treatment from the hospital and patient’s perspective for hospitalized and ambulatory case. A total of 311 patients were enrolled in this study. Of these 239 cases were inpatients while 72 cases were outpatients. The average treatment cost of a hospitalized rotavirus episode was US$ 667.76 and out of this US$ 480.36 (72%) was by the hospital, while US$ 187.40 (28%) by the patient. The average cost of treatment for ambulatory rotavirus case was US$ 166.34, of which US$ 120.02 (72%) was from hospital cost and US$ 46.32 (28%) from patient’s cost. These costs for inpatient and outpatient were during the whole treatment period. Hospital and patient’s cost of rotavirus treatment showed the disease posed a considerable economic burden on Libyan government and families. Applying preventive measures would be more appropriate such as introducing rotavirus immunization to reduce the disease.