BACKGROUND Different methods are available for diagnosis of visceral leishmaniasis (VL), among them the urine-based antigen detection assay, latex agglutination test (Katex), is a recently developed one. The main drawback of the test is false-positive reactivity in some of healthy individuals. The false positivity of the test can be removed by boiling the urine sample for 5 min before testing. In this study an attempt was made to improve Katex by removing unpleasant boiling process, which also decreases field applicability of the test. METHODS False-positive and true-positive urine samples were collected from VL patients and healthy individuals. Both samples were then treated by reagents including, sodium dodecyl sulfate, trichloroacetic acid, dithiothreitol (DTT), sulphosalicylic acid and also heating at 56 degrees C. RESULTS Findings of this study showed that DTT pretreatment significantly reduced the rate of false-positive reactivity of Katex where 73% of false-positive urine samples changed to negative after DTT treatment. However, the DTT treatment reduced the rate of true positivity by 14%. CONCLUSIONS These data indicate that DTT can be used to eliminate nonspecific reactivity in the Katex. This will improve the performance of Katex and make it a more convenient and field applicable test.