Scrub typhus is a zoonotic bacterial disease caused by Orientia tsutsugamushi (Rickettsiales: Rickettsiaceae) and trombiculid mite larvae, also known as chigger, are known vector. Until recently, O. tsutsugamushi is the only species of the genus. However, two new species Candidatus O. chuto and Candidatus O. chiloensis that causes scrub typhus were reported recently in the Middle East, southern Chile, and Africa. In addition, Orientia spp. bacteria detected from field collected free-living Eutrombicula chigger mites in the United States. Despite these trends, research on new species is insufficient in the Republic of Korea. Therefore, we focused on finding the presence of novel species or strains from chiggers. Specimen chiggers harvested from rodents collected in 16 regions in October 2022 collaborating with the Regional Center for Vector Surveillance against Climate Change were selected. A total of 1,249 specimen belonging 4 genera and 14 species were identified by using a fluorescence microscope and 266 pools were produced by pooling up to 10 individuals per species chiggers. To detect Orientia spp., we screened by using a real-time PCR targeting the 16S rRNA gene. Overall minimum infection rate was 0.56% (7 pools/1,249 tested specimen). After screening, conventional nested-PCR for targeting 47-kDa htrA gene was conducted to obtain sequences, and four of the positive pools were amplified. Through phylogenetic analysis, three pools were clustered with O. tsutsugamushi Gilliam and UT221 strain, and the other pool was formed a clade that was distinct from O. tsutsugamushi. These results suggest that novel species of Orientia may exist in the Republic of Korea.