Lawsonia intracellularis is a bacterium already described in several species and most prevalent in pigs, in which it causes enteric problems. Horses can also be affected, developing a disease known as equine proliferative enteropathy, which results from the proliferation of intestinal crypt cells in response to infection by the bacterium. Despite the existence of reports of the disease in several countries, including Brazil, there are still no reports of the disease or epidemiological studies of its occurrence in symptomatic or asymptomatic horses in the state of Paraná. Thus, the present study was conducted to examine the occurrence of L. intracellularis in asymptomatic horses raised in the west, northwest and north regions of Paraná by means of serological testing and the real-time polymerase chain reaction (qPCR) technique. In the serological approach, the immunoperoxidase monolayer assay (IPMA) technique was employed. Feces were processed and subjected to qPCR. In total, samples were collected from 162 animals from 20 farms. Of these, 9/162 (5.55%) showed specific antibodies against L. intracellularis. Real-time PCR, on the other hand, identified 7/162 (4.32%) fecal samples positive for the presence of the bacterium. When the techniques were compared, none of the samples was positive by both, demonstrating that, for a better diagnosis, they must be performed together. In contrast to most reports in horses, the present study describes higher serological and molecular occurrence in animals older than two years. These results are of great epidemiological relevance, as they indicate that the bacterium is present in the sampled regions of the state of Paraná. Therefore, the disease must be included in the differential diagnosis of diseases with similar clinical manifestations.