BackgroundScrub typhus and murine typhus, caused by Orientia tsutsugamushi and Rickettsia typhi, respectively, are important causes of febrile illness in Laos. Although several studies have assessed rickettsial infection in selected provinces, the nationwide distribution remains unclear. This study aimed to estimate exposure to scrub typhus group (STG) and typhus group (TG) across twelve provinces of Laos and identify potential hotspots.MethodologyWe screened 1,200 serum samples from blood donors (100 per province) for STG and TG IgG antibodies using enzyme-linked immunosorbent assays (ELISA). Samples with optical density ≥ 0.5 were confirmed by immunofluorescence assays (IFA).ResultsOverall seroprevalence was 7.26% (95%CI:5.93-8.87) for STG and 4.09% (95% CI:3.11-5.37) for TG. STG seroprevalence was highest in Huaphan (27%), Oudomxay (19%) and Xiangkhuang (17%), all in northern Laos. TG seroprevalence was 10% in both Oudomxay (north) and Attapue (south). Compared with Vientiane Capital, these provinces had significantly higher seropositivity. No significant association was observed with age group. STG seropositivity was higher in males, whereas TG seropositivity was higher in females.ConclusionRickettsial infections are widespread in Laos, with STG seroprevalence concentrated in the north and TG seroprevalence present in both the north and south, guiding future research priorities and informing targeted public health interventions.