SummaryTwo batches ofCicadulina mbilawere given two distinct acquisition access periods (AAP) (3 h and 50 h) on maize plants infected with maize streak virus (MSV). Infectivity assays on susceptible maize were carried out 1, 3, 10, 17, 26 and 35 days after the AAP. Transmission efficiency was significantly higher forC. mbilasubjected to the 50‐h AAP. At the same time as the infectivity assays, the amount of MSV in each leafhopper was determined by an indirect double antibody sandwich (IDAS) ELISA. There were more ELISA‐positive insects after the 50‐h AAP than after the 3‐h AAP. In the group given a 3‐h AAP, only 7% of the insects tested between day 1 and 35 were found to be positive by ELISA. In contrast, after the 50‐h AAP, the majority ofC. mbilawere positive, yet a decrease in ELISA‐positive insects was noticed from day 17 onwards. Using a calibration curve obtained with purified virus, as little as 0.15 ng of MSV per insect could be measured by the IDAS‐ELISA. A mean value of 0.36 ng of MSV perC. mbilawas found 3 days after the 50–h acquisition, whereas 14 days later there was only 0.20 ng of virus per insect. For comparison, when leafhoppers were kept on infected maize, they displayed substantial accumulation of MSV up to an average of 3.83 ng of MSV per insect after 35 days of continuous acquisition. The amount of virus per insect detected in females was usually greater than the amount detected in males. Our results suggest that MSV does not multiply inC. mbilaand contribute to the understanding of the persistence of transmission efficiency in the absence of vir