Chigger mites have a cosmopolitan distribution and represent an important public health concern due to their ability to cause dermatitis and to transmit pathogens associated with disease. Their larval stage parasitizes all terrestrial vertebrates including humans, who serve as accidental hosts. Research on trombiculid mites expanded substantially during the 1940s and 1950s, particularly throughout World War II, leading to a marked increase in the number of described species. For many years, the genus Orientia was considered monotypic, comprising only Orientia tsutsugamushi, with its distribution thought to be restricted to the so-called “Tsutsugamushi Triangle.” This information was challenged by reports of scrub typhus cases in southern Chile, in Chiloé Island. Following these initial detections, it was assumed that Orientia circulation was limited to the Chiloé region. Therefore, the objective of this study was to conduct a comprehensive Orientia surveillance in mites collected across the Chilean territory. Mite samples used in this study were collected from wild rodents across Chile, encompassing the Big North, Small North, Central, and Southern regions. The samples were processed either individually or in pools of up to 40 mites originating from the same host. Genomic DNA was extracted from ethanol-preserved specimens using the DNeasy Blood & Tissue Kit (Qiagen). Conventional PCR assays targeting endogenous mite genes were first performed to assess sample integrity. Samples that tested positive for one or both endogenous markers were considered viable and were subsequently screened by qPCR for bacteria of the genus Orientia, using genus- specific primers targeting the 16S rDNA gene. A total of 126 samples were evaluated, of which 75 were analyzed individually and 51 as mite pools. Voucher specimens were slide-mounted and morphologically identified as Carebareia sp. Goff and Brennan, 1977; Morelacarus sp. Vercammen-Grandjean, 1973; Paratrombicula chilensis Stekolnikov and González-Acuña, 2012; Quadraseta chiloensis Stekolnikov and Silva-de la Fuente, 2020; and Quadraseta sp. Brennan, 1970. Among the tested samples, 19 were positive for Orientia. These detections originated from mites identified as Quadraseta sp. and Carebareia sp. Notably, this is the first report of Orientia DNA in a mite of the genus Carebareia. One of the positive samples corresponded to a protonymph originally collected as a larva in Cochamó, Los Lagos, and reared under laboratory conditions until the protonymph stage. This finding demonstrates that chigger mites are capable of maintaining Orientia across transstadial developmental stages.