In this study we have evaluated four fluorescent antibody reagents, three monoclonal and one polyclonal, for identification of adenovirus isolates and compared four conventional cultures (human embryonic kidney, A549, HEp-2, and MRC-5 cells) with centrifugation culture for rapid diagnosis. For identification of adenovirus isolates by immunofluorescence, CDC reagent and Adenoclone, both monoclonal antibodies to the hexon group-reactive antigen, were more sensitive and easier to interpret than the other two reagents tested. HEK and A549 cells were the most sensitive for isolation of adenovirus. Although A549 cells were an inexpensive alternative to HEK, A549 cell monolayers deteriorated more rapidly and passages were more often required. Centrifugation cultures with A549 cells detected 77% of positives within 2 days and 100% within 5 days, whereas isolation in conventional culture required up to 10 days for HEK and up to 20 days for MRC-5 cells.