A new direct agglutination test (DAT) for use with urine samples for the diagnosis of visceral leishmaniasis (VL) has been developed and compared with the conventional DAT with serum samples and our previously reported enzyme-linked immunosorbent assay (ELISA) with urine samples (urine ELISA). The new DAT, in which anti-human IgG was used as enhancing antibody, was tested with urine samples from 75 VL patients and 225 non-VL patients and healthy people. The sensitivity of the new DAT (90.7%), was almost the same as that of the conventional DAT (91.0%) and the urine ELISA (93.3%). The specificity of the new DAT (96.4%) was nearly identical with that of the urine ELISA (97.3%). A urine-based DAT has several advantages over the conventional DAT: sample collection is non-invasive and it can process larger numbers of samples with smaller amounts of antigen.