An available enzyme-linked immunosorbent assay (ELISA) was studied for the detection of anti-Dioctophyma renale antibodies in the sera of dogs using, detection of parasite eggs in urine sediment as a reference test. ELISA uses a soluble antigenic preparation of esophagus of D. renale and the optimal dilutions of the antigen, serum and conjugate were determined by means of checker board titration, using positive (n=13) and negative (n=27) reference serum. The specificity and sensitivity of the ELISA were 93.8% and 92.3% respectively and the kappa index was good (0.76). These results suggest that ELISA described may prove to be an effective serological test for detecting dogs infected and exposed to this parasite mainly dogs that are not eliminating parasite eggs through their urine.Department of Veterinary Medicine, Universidade do Contestado (UnC), Rua Roberto Ehlke 86, CEP 89460-000 Canoinhas, SC, BrazilDepartment of Preventive Veterinary Medicine and Animal Reproduction, School of Agrarian and Veterinary Sciences (FCAV), Universidade Estadual Paulista (UNESP), Via de Acesso Prof. Paulo Donato Castellane s/n, CEP 14887-900, Jaboticabal, SP, BrazilDepartment of Veterinary Pathology, FCAV/UNESP, Via de Acesso Prof. Paulo Donato Castellane s/n, CEP 14887-900 Jaboticabal, SP, Brazil.Department of Preventive Veterinary Medicine and Animal Reproduction, School of Agrarian and Veterinary Sciences (FCAV), Universidade Estadual Paulista (UNESP), Via de Acesso Prof. Paulo Donato Castellane s/n, CEP 14887-900, Jaboticabal, SP, BrazilDepartment of Veterinary Pathology, FCAV/UNESP, Via de Acesso Prof. Paulo Donato Castellane s/n, CEP 14887-900 Jaboticabal, SP, Brazil.