Several Plasmodium falciparum asexual stage antigens have been reported as possible candidate antigens for vaccines against malaria. One such is the 96-kDa antigen. We used the 96 tR recombinant protein to determine the levels of antibodies to this protein over a 6-mo period in children 4 mo to 15 yr old, who make up the population at risk in Cameroon. The mean enzyme-linked immunosorbent assay (ELISA) on 83 children at the start of the study in June was 0.270, with 13% of these children having ELISA values considered to be positive (> or = 0.36). In the follow-up study in early December, the mean ELISA was 0.320 with 20% of the children having positive values. In the period between June and December, 85% of the children who returned for follow-up had had 1 or more attacks of malaria. The mean ELISA values of this latter group had increased from 0.235 in June to 0.318 in December, with 6% of the children in June having positive values compared with 18% in December. These findings reinforce our evidence from a previous study that reactivity to the 96-kDa antigen is related to adequate exposure or repeated infection.