To measure protection induced by natural rotavirus infection, 163 infants enrolled in a rotavirus vaccine trial were prospectively followed for 2 years. Serotype 1 rotaviruses were the predominant circulating strains during the study. Over the 2 years of observation, significantly fewer infants infected before enrollment developed a symptomatic reinfection (0 of 21) or any reinfection (4 of 21) compared with previously uninfected infants (P = .0003). Of the 60 infants who developed a primary rotavirus infection in the first year (40 symptomatic, 20 asymptomatic) only 4 were reinfected in the second year compared with 29 of 82 subjects not previously infected (P = .00003). Asymptomatic primary infection appeared to be as protective as symptomatic primary infection. The only symptomatic reinfections occurred in 2 subjects who did not develop rotavirus antibody after the initial detection of rotavirus. An age-related reduction in the ratio of symptomatic to asymptomatic primary rotavirus infection was also detected. In this study, protection against homotypic serotype 1 reinfection appeared to last greater than or equal to 2 years.