Objective To investigate the infection status of Toxoplasma gondii in different colonies of tree shrews and then provide the basis for parasitological monitoring. Methods Each of the forty blood samples were randomly collected from three tree shrews colonies: wild origin, domesticated and first generation, respectively. Both indirect hemagglutination test (IHA) and PCR assay were used to detect the Toxoplasma gondii. Results No positive sample of Toxoplasma gondii was detected from either IHA or PCR results. The results from IHA and PCR assays were in coincidence with each other. Conclusions According to the survey none of the tree shrews from the three groups is infected with Toxoplasma gondii. More samples or infection experiments are needed to determine whether tree shrews can be infected with Toxoplasma gondii.