A sensitive and specific 30-min Dot-ELISA for the detection of anti-Leishmania antibodies in the dog

A very rapid, sensitive and specific Dot-ELISA was developed for diagnosing visceral leishmaniosis in dogs infected with Leishmania infantum. Sera from 26 healthy dogs and 127 dogs with different infectious diseases including 40 dogs with leishmaniosis, and 87 dogs with other suspected or confirmed infectious diseases, was evaluated in an alkaline phosphatase ELISA on a nitrocellulose membrane, sensitized with soluble promastigote antigen. The test procedure lasted only 30 min. Distinction between leishmania positive and leishmania negative sera was complete at a dilution of 1/320, hence there was no cross-reactivity, not even with sera from dogs with trypanosomal or babesial infections. Therefore, it is concluded that this test has a very high sensitivity and specificity for the detection of anti-leishmania antibodies in the dog. The Dot-ELISA was significantly positive correlated with the direct agglutination test (DAT), the indirect immunofluorescence test (IFAT), and the Slide-ELISA. A significant concordance of the four tests was also demonstrated.