Lipophosphoglycan (LPG) is the major surface glycoconjugate of Leishmania protozoan and has an important biological role in host-parasite interactions both in the midgut epithelium of the sand fly vector and in the vertebrate macrophages. Canine leishmaniasis (CanL) is a chronic infectious disease predominantly caused by Leishmania infantum. An early and accurate immunodiagnosis of the disease is crucial for veterinary clinical practice and for disease control. In this work, we evaluated L. infantum LPG as an antigen in an indirect enzyme-linked immunosorbent assay (ELISA) for CanL immunodiagnosis (LPG-ELISA) by testing serum samples from 97 naturally infected dogs with diverse clinical presentations ranging from subclinical infection to severe disease, as evaluated by veterinarian infectologists. Serum samples from healthy dogs from non-endemic areas (n = 68) and from dogs with other infectious diseases (n = 64) were used as controls for assay validation. The performance of the LPG-ELISA was compared with that of an ELISA using the soluble fraction of L. infantum total lysate antigen (TLA). LPG-ELISA presented a superior performance in comparison to TLA-ELISA, with 91.5% sensitivity, 98.5% specificity and 99.7% accuracy. A distinguishing feature of the LPG-ELISA compared to the TLA-ELISA was its higher ability to identify subclinical infection in clinically healthy dogs, in addition to the absence of cross-reactivity with other canine infectious diseases. Finally, LPG-ELISA was compared to TR DPP visceral canine leishmaniasis test, the immunochromatographic test recommended by the Brazilian Ministry of Agriculture. LPG-ELISA exhibited higher values of specificity (98.5% versus 93.1%) and sensitivity (91.5% versus 90.6%) compared to TR DPP. In conclusion, L. infantum-derived LPG was recognized by antibodies elicited during CanL in different infection stages and was shown to be a suitable antigen for specific clinical settings of veterinary diagnosis and for public health usage. Canine leishmaniasis (CanL) caused by Leishmania infantum is a zoonotic disease with high importance for the public health of several countries. L. infantum-infected dogs can be a domestic reservoir of the protozoan parasite for sand flies, which transmit it from dogs to humans during their blood meals. It is important to diagnose and treat infected dogs as early as possible, so the dissemination of CanL and the transmission to humans can be controlled. The currently commercially available assays present problems, such as cross-reaction with other canine diseases and lack of sensitivity in the detection of dogs that do not present clinical signs of disease. In this work, we tested lipophosphoglycan (LPG), a molecule abundantly found in the protozoan ' s surface, as an antigen in an immunodiagnostic platform. We were able to see that the assay using LPG is highly sensitive and specific, showed no cross-reaction with other canine infectious diseases and successfully identified infected dogs with no signs of disease.