Polymerase chain reaction (PCR) and restriction endonuclease fragment length polymorphism (RFLP) analysis were used to characterize the genotypic diversity of three isolates of spotted fever group (SFG) rickettsiae isolated from ticks in China. A primer pair designed from DNA sequence encoding 190 K protein antigen of R. rickettsii and genomic DNAs obtained from the isolates were used in PCR. The PCR products were cleaved with restriction endonucleases PstI and RsaI, and the digestion patterns were analyzed by polyacrylamide gel electrophoresis (PAGE) and compared with those of all known species and strains of SFG rickettsiae. The results showed that three isolates had the same PCR products as the other SFG rickettsiae under comparison. HL-93 strain, isolated from Hemophysalis concinna ticks collected in Hulin County, Heilongjiang Province, had unique PstI digestion pattern among SFG rickettsiae; strains BJ-93 and 053, isolated from Dermacentor sinicus and Haemaphysalis concinna ticks collected in Changping County, Beijing City, and Suifenhe City, Heilongjiang Province, respectively, had the same PstI and RsaI digestion patterns as strains R. sibirica 246, BJ-90 and IMTO-85. The present study demonstrated that the BJ-93 and 053 strains were genotypically identical with R. sibirica and the HL-93 strain was genotypically unique among SFG rickettsiae.