Abstract: Objective To identify the pathogen of patients with fever with unknown origin and determine whether they were infected with spotted fever group Rickettsia. Methods Nested polymerase chain reaction (nPCR) was conducted to amplify groEL gene of rickettsia from the blood samples of the patients, the nucleotide sequencing and identification were carried out. Results Rickettsial DNA was detected in 1 of the 20 patients (5%). The groEL gene of the strain shared high homology (99%) with Rickcttsia rickcttsii Iowa, Rickcttsia peacockii Rustic and Rickcttsia sibirica. Conclusion It was the first time to diagnose the patient with spotted fever in Dali, Yunnan province. It is necessary to enhance the differential diagnosis of spotted fever in local medical institutions.