Abstract: DF1 cells were inoculated with three different doses (100, 10, 1 μL) of ALVA, ALVC, ALVJPY and ALVJWS separated from different chickens to evaluate three kinds of A, B, C ELISA kits. The samples of DF1 cells supernatant were obtained at different days post inoculation (dpi) and detected with three kinds of kits after freezing and thawing once. The results indicated that the samples inoculated with ALVA or ALVJPY were both detected positively firstly at the 3rd dpi with A ELISA kit for 100 μL inoculation doses and at the 7th dpi for 1 μL dose; If with B ELISA kit, two ALVs of subgroup were detected positively within some days post inoculation, but the detectable time were longer than that of A ELISA kit; If with C ELISA kit, the detectable time of viruses were the longest. ALVC and ALVJWS were detected positively with A ELISA kit at the 5th and 9th dpi for 100 μL doses, respectively; but for the other doses, no positive samples were detected at whole observed days. If with B and C ELISA kits, none of inoculated cells with three doses of inoculation was detected positively. It can be concluded that it is dramatically different between A, B and C ELISA kits in the detection of three exosomic subgroups of ALVs；The A ELISA kit is more sensitive than other two ELISA kits (B and C). Simultaneously, the positively detected days to ALVs are related to the doses of inoculation in DF1 cells. The results can be helpful for the application of three ELISA kits to the detection or isolation of exosomic ALVS.