In a phase 1 study to evaluate human-rhesus rotavirus reassortant vaccines, 116 infants 1 to 5 months of age received one of the following five preparations: the serotype 1 reassortant, the serotype 2 reassortant, rhesus rotavirus (serotype 3), a bivalent preparation (serotypes 1 and 3), or a placebo. Seroresponses to the different vaccines were measured by plaque reduction neutralization assay (PRNA); rotavirus-specific immunoglobulin A (IgA), IgG, and IgM enzyme-linked immunosorbent assays (ELISAs); and complement fixation (CF). The seroresponse rate, calculated by using a fourfold or greater antibody rise by any assay, was similar in the four vaccine groups (83 to 96%). When the data from all the vaccinees were pooled, IgA ELISA, IgG ELISA, and PRNA were comparable in detecting seroresponses (67, 62, and 70%, respectively) and more efficient than IgM ELISA (53%) and CF (44%). When the vaccinees were analyzed by age, the overall seroresponse rates were the same for infants 1 to 2 and 3 to 5 months old (90%). The IgA ELISA and PRNA were the most efficient for detecting antibody rises in both age groups. IgG ELISA was among the least efficient methods for detecting antibody rises in the younger age group but among the most efficient in the older age group (44 versus 78%). CF was among the least efficient methods in both age groups but was significantly better in the older age group than in the younger age group (54 versus 21%). Our findings show that ELISA, in particular rotavirus-specific IgA ELISA, is a sensitive indicator of vaccine takes in 1- to 5 month-old infants, the target population for vaccination. ELISA should also be very useful in demonstrating natural rotavirus infections in field studies in which a stool specimen from a diarrheal episode is not always available. The ELISA has the advantages of being easier and quicker and requiring less serum than PRNA, but it does not give serotype-specific information about the immune response.