A lateral flow immunoassay based on colored latex particles for detection of canine visceral leishmaniasis

Abstract

Canine visceral leishmaniasis (CVL) is the major source of human visceral leishmaniasis. To control the spread of this disease, early and accurate detection of infected dogs is critical but challenging. The serological diagnosis of CVL remains problematic because there are no reliable commercially available tests. Most laboratories use enzyme-linked immunosorbent assay or the indirect immunofluorescent antibody test. These tests use Leishmania chagasi recombinant antigens K39 or K26 assembled with either gold-labelled Staphylococcus aureus protein A or protein G from Streptococcus pyogenes. In this work, we propose the development, optimization and standardization of a lateral flow immunoassay (LFIA) based on functionalized colored particles and a specific recombinant antigen, as a visual in situ method for the diagnosis of CVL. The following analysis variables were considered: i) the concentration of the latex-protein complex; ii) the dilution of the serum; iii) the composition of the employed buffers; iv) the nominal capillary flow time through the nitrocellulose membrane; v) the concentration of reagents fixed in the test and control lines; vi) the particle size of the colored latex; and vii) the conjugation method. Then, the obtained strips were evaluated as a visual diagnostic tool based on a panel of positive and negative sera. It was observed that because of its simplicity and performance the LFIA test is a quick and reliable alternative for the diagnosis of CVL either in conventional laboratories or for remote areas where laboratories are not readily accessible for conventional assays.