Abstract: Objective To investigate the natural infection status of Rickettsiae tsutsugamushi in rodents in Lishui, and provide scientific evidence for the prevention and control of the disease. Methods The liver and spleen specimens of the rodents captured in Lishui were collected to detect the 56 kD gene by nested polymerase chain reaction (nPCR). Results Totally 132 rodents were captured in Lishui. Three specimens of Apodemus agrarius and 1 specimen of Mus musculus were positive for Rickettsiae tsutsugamushi. The positive rate was 3.03% (4/132). Conclusion The finding indicated that Rickettsiae tsutsugamushi infection do exists in rodents in Lishui and the host animals are diversified, which could be used in the further investigation of tsutsugamushi focus in Lishui.