To screen single cell clone line for cultivation of PCV2 with the highest titer, dilution method was used to isolated single clone lines from PK15 heterogeneous cells. The obtained single cell clones were subjected to PCV2 infection through synchronized inoculation at a ratio of 1∶100. PCR and quantitative PCR analysis showed the yield of PCV2 in cell clone A10 was the highest after PCV2 infection. Then second cell clone was conducted on A10 cell to ensure the homogeneous character of A10, followed by PCV2 inoculation test through synchronized and non-synchronized inoculations. The result of quantitative PCR indicated the maximum PCV2 genomic copy numbers obtained with synchronized and non-synchronized inoculations were 3.0×109/mL and 6.2×109/mL, respectively, which were higher than the maximum PCV2 genomic copy numbers with PK15 heterogeneous cells. These results suggest the constructed homogeneous PK15 cell line is more suitable to cultivate PCV2 compared to PK15 heterogeneous cells.