ABSTRACT. Lawsonia intracellularis is an obligate intracellular pathogenic bacterium that causes proliferative enteropathy in domestic and experimental animals. Antiserum against synthetic peptides of the Lawsonia surface antigen (LsaA) well recognized L. intracellularis in infected ileum by immunohistochemistry. The synthetic peptides in LsaA showed strong reaction with serum from rabbits infected with L. intracellularis by enzyme-linked immunosorbent assay. These results suggest that ELISA used synthetic peptides in LsaA and anti-LsaA serum might be useful to diagnose for proliferative enteropathy. KEY WORDS: ELISA, Lawsonia intracellularis, rabbit.J. Vet. Med. Sci. 66(6): 735-737, 2004 Proliferative enteropathy (PE) is an intestinal infectious disease characterized by thickening of the aboral small and proximal large intestinal mucosa due to enterocyte proliferation associated with the presence of an intracellular bacterium [11]. This obligate intracellular bacterium, the causative agent of PE, was characterized as a novel genus and species and was named Lawsonia intracellularis [10]. L. intracellularis has been associated with colonization of enterocytes in rabbits, hamsters, rats, guinea pigs, swine, sheep, horses, white-tailed deer, dogs, arctic fox, ferrets, and ostrich [1,5,12]. Because of the development of L. intracellularis specific primers [6], detection of fecal shedding by PCR has been possible. Serum IgG specific against L. intracellularis has been detected in pigs by an immunofluorescence antibody test [4,7] or immunoperoxidase monolayer assay [2,3]. However, serological detection of L. intracellularis is not established in Japan. In this study, we report a case of a rabbit infected with L. intracellularis in Japan and detection of the bacteria by antiserum for Lawsonia surface antigen.A six-week-old female New Zealand White rabbit showed diarrhea, and fecal samples were collected and tested for L. intracellularis DNA by PCR [6] to check the infection of L. intracellularis. As the fecal samples were positive by PCR, the rabbit was suspected of being infected with L. intracellularis. Thickening of the gut wall was also observed. To confirm the infection of L. intracellularis, we cultivated L. intracellularis by cell culture method [8]. Frozen (-80°C) intestine was homogenized and was used as the source material. Rat small intestinal cells (IEC-18; ATCC CRL 1589) were grown in Dulbecco's modified Eagle medium (DMEM; Sigma) with 10% fetal calf serum (FCS) on glass coverslips on a 6-well plate. Monolayers were exposed to the source material and then were centrifuged at 700 × g before transfer to an incubator. The incubator was kept at an atmosphere of 8.0% O 2 and 8.8% CO 2 at 37°C.The culture was incubated for 6 hr and the medium was replaced with DMEM containing gentamicin (30 µg/ml), and then was transferred to the incubator. At 6 days after infection, the infected monolayer examined by phase microscopy showed little morphological change and several detached cells (Fig. 1B). To detect intracell...