Transmission of Plasmodium falciparum from man to mosquito can be affected by human sera. Whereas serum-dependent reduction of transmission has been shown to be reproducible, there is limited evidence for enhancement of transmission. We aimed to assess the prevalence and reproducibility of transmission enhancement (TE) by human sera from different geographic areas (n = 642), in comparison with the capacity for transmission reduction (TR). The overall prevalence of TE (7%) was lower than that of TR (48%) and its effect generally weaker but reproducible in repeated measurements. TR but not TE showed a significant association with the presence of serum antibodies against Pfs48/45 and a non-significant trend to the presence of anti-Pfs230 antibodies.