Objective To Study of in situ polymerase chain reaction assay for the detection of canine parvovirus(CPV) in Feline Kidney F 81 cells.Method Feline kidney F 81 cells were cultured in slide and infected with CPV. The cells infected with CPV in different stages were detected by direct in situ PCR, which used biotin labelled primers, and immunohistochemical method. Result In situ PCR method, a lot of signals were found in cells infected from 12 to 48 hours. Some signals were found in cytoplasm, and some in nucleus. In immunohistochemical method, no signals were found in cells infected for 12 hours, and sporadic signals in cells infected for 48 hours. The signals detected by these two methods showed significant difference. Conclusion The results indicate that the in situ PCR not only has high sensitivity, but also can locate the position of virus.