Validation of Enzyme-Linked Immunosorbent Assay (ELISA) with recombinant antigens for the serological diagnosis of canine visceral leishmaniasis (VL)

Abstract

In Brazil, visceral leishmaniasis (VL) is booming and present in all regions, with a prevalence of 3,253 cases in 2013, reaching also urban and peri-urban centers in the last three decades. It is caused by Leishmania (Leishmania) infantum. Sandfly Lutzomyia longipalpis is the main vector, and the main reservoirs, involved in domestic and wild cycle, respectively, are the dog (Canis familiaris) and fox (Dusycion vetulus). As dogs have an intense parasitism in the skin, it is believed that this will allow an easy vector infection constituting the important link in maintaining the epidemiological chain. Thus, the identification of infected dogs is considered a crucial measure in controlling the transmission of the disease. It is recommended to perform sample or census survey, depending on the epidemiological situation regarding the risk of transmission in the area being investigated. The Ministry of Health recommends conducting immunoassay DPP " Leishmania test for the screening of dogs and ELISA based on total lysate of Leishmania major-like for the confirmation of positive cases. Since the parasite culture for preparation of the total antigen constitutes a limiting factor, it would be desirable to have an ELISA based on the use of recombinant antigen, which we propose in this project. Thus, this study aims to validate the serological methods ELISA-rHsp83 and ELISA-rK28 for the diagnosis of canine VL replacing ELISA based on total lysate of Leishmania major-like as a confirmatory test after immunochromatographic DPP " Leishmania test. Validate alternatively serological methods ELISA-rHsp83 and ELISA-rK28 for the diagnosis of canine VL as a confirmatory test, regardless of the outcome of DPP " Leishmania test. (AU)