We evaluated the cost-effectiveness of rotavirus vaccination in Uzbekistan from the healthcare system and societal perspectives. Disease burden was estimated using national statistics on hospitalizations and deaths, and international estimates of under-five mortality. Without vaccination, the risk for rotavirus hospitalization by age 5 is 10 per 1000 children. Rotavirus hospitalizations cost US$ 406,000 annually, of which US$ 360,000 (89%) is for medical expenses and US$ 46,000 (11%) is for non-medical and indirect costs. Rotavirus mortality rate at 0.7 per 1000 derived from national data was three-fold lower than the same rate calculated from international estimates of under-five mortality. Rotavirus vaccination could reduce hospitalizations and deaths by 91% and avert US$ 370,000 in hospitalization costs alone. Vaccination would be cost-effective with vaccine prices in a range of US$ 2-25 per child. However, the cost-effectiveness is greatly influenced by mortality, vaccine price and vaccine efficacy.