Rotavirus (RV) is the most common cause of severe gastroenteritis in infants and young children worldwide. RV causes approximately half a million deaths each year among children aged <5 years. According to WHO estimates for 2008, there were approximately 10 to 50 deaths annually in young children due to rotavirus diarrhea in Georgia. The purpose of this study was to assess the knowledge, attitudes, and practices related to rotavirus diarrhea and the rotavirus vaccine among health care workers (HCWs). The National Center for Disease Control and Public Health (NCDC) conducted a cross-sectional survey of HCWs involved in the expanded program of immunization (EPI). The HCWs were selected by simple random sampling using employee lists, and questionnaires were hand-delivered to selected HCWs during working hours. The majority of HCWs (67.9%) recognized diarrhea as a frequent health-related problem in children under 2 years of age. However, 53.9% partially disagreed with the statement that rotavirus is the most common cause of all forms of diarrhea. Multivariable analysis revealed that the following perceptions among HCWs more than doubled the likelihood that they would not support adoption of the RV vaccine: no perception of need for this vaccine in Georgia specifically (OR=2.54, CI: 1.46-4.42), no perception of need to address burden of disease in developing countries (OR=2.75, CI: 1.59-4.75), and concerns about the vaccine's safety (OR=2.51, CI: 1.56-4.02). Concerns about vaccine safety, low awareness about the RV disease burden and the effectiveness of the RV vaccine, combined with a lack of communication skills represented the main predictors of RV vaccine uptake among HCWs. Intensive vaccine promotion interventions that focus on the epidemiology of disease and vaccine effectiveness are urgently needed.