Objective To analyze the tissue distribution, viral excretion and serological antibody in BALB/c mice artificially infected with minute virus of mice (MVM), and the natural infection status in mice. Methods Thirty-three SPF male 3-week old BALB/c mice were intraperitoneally injected with 0.2 mL of MVM in 1.2×10⁷ copies/μL concentration. The general status of the animals was observed daily post inoculation. The animals' tissue,faeces and serum samples were taken at 12 time points before and after inoculation (2-3 animals at each time point). QPCR method was used to detect the viral nucleic acid in tissues and feces, and the serological antibody against MVM was tested by ELISA. Meanwhile,other clinical samples of 1563 SPF mice and 158 mice in open housing were collected for testing the viral nucleic acid and antibody. Result There were no clinical symptoms and pathological changes among all infected mice. The viral loads of each tissue reached the peak at 4 d or 7 d post inoculation, and then were generally on a declining curve, but were still found at 60 d. Comparing the viral load in tissues showed that the highest tissue was liver, followed by kidney, spleen, stomach, heart, lung, cecum and brain. The viral loads in feces reached the peak at 11 d, and then dropped rapidly, but could be still detected at 60 d. The antibody could be detected at 7 d, and then gradually raised. The antibody dilution degrees reached 32 at 21 d, and maintained high levels with 16 to 128 during 32 d to 60 d. In the clinical samples, the nucleic acid tests were negative in SPF mice, however, a positive rate of 14.5% was found in mice in open housing. Meanwhile, the antibodies of MVM showed a low positive rate in SPF mice, and the positive rate was 68.3% in the mice in open housing. Conclusions The mice generally present occult infection after inoculation of MVM. However, the infected mice can shed virus in a long period, and viral loads of tissues and serological antibody can be maintained for a long time. Thus, MVM can be detected by testing viral nucleic acid and serological antibody.