A high titer of hemagglutinin (HAnin) was found in culture fluids of cells of Aedes albopictus, clone C6/36, infected with type 1 dengue virus. The HAnin (TC antigen) was associated with complete virions and no appreciable small-sized HAnin was produced, in contrast to the case in an infected suckling mouse brain (SMB) homogenate, in which most of the HAnin is smaller than complete virions. Extraction of the TC antigen with Tween 80-ether disrupted the HAnin into smaller-sized particles, resulting in complete loss of infectivity (TE-TC antigen), but it did not affect the titer or reactivity of the HAnin. Comparative hemagglutination-inhibition (HI) assay of human sera using TC antigen, TE-TC antigen or standard antigen extracted from infected SMB (SMB antigen) showed that TC or TE-TC antigen could be used for routine diagnostic or epidemiological HI tests instead of SMB antigen, which is rather hard to prepare.