Rotavirus gastroenteritis is a disease that causes significant morbidity and mortality worldwide in infants and children. The recent availability of efficacious rotavirus vaccines with favorable safety profiles has prompted medical agencies around the world to register the vaccines and countries and regions to recommend universal vaccination of healthy infants to prevent the serious health and economic consequences of rotavirus illness in children. Premature infants can be especially vulnerable to severe rotavirus infections, which may lead to serious outcomes. The clinical efficacy and safety of the pentavalent human-bovine rotavirus vaccine (PRV, RotaTeq, Merck, Whitehouse Station, NJ, USA) in healthy infants has been evaluated in the large phase III Rotavirus Efficacy and Safety Trial (REST). Preterm (N = 2,070) and exclusively breastfed infants (N = 1,566) were included in this study. The efficacy and safety of PRV in premature infants and in breastfed infants from the REST database are reviewed along with guidelines from national medical organizations for the use of PRV in these populations. Based on these data from the REST study, premature infants receive the same protection from PRV compared with nonpremature infants, without additional safety risk. Similarly, breastfed and nonbreastfed infants are equally protected from severe consequences of rotavirus gastroenteritis.