Testing different antigen capture ELISA formats for detection of Leptospira spp. in human blood serum

Abstract 

Leptospirosis is an infectious disease caused by pathogenic spirochetes of the genus Leptospira. The illness is characterized by an acute bacteremic phase followed by an immune phase, in which specific antibodies are found in blood and leptospires are eliminated in urine. Novel diagnostic strategies for use in the acute phase of leptospirosis are needed since clinical manifestations in this phase mimic other feverish tropical diseases. In the present study, mAbs and polyclonal IgY were used in the standardization of three different antigen capture ELISA formats for direct detection of leptospires in human blood during the acute phase of the disease. Detection limit of leptospires in experimentally contaminated human sera ranged from 10⁵ to 10⁷ cells ml⁻¹ in the different formats. The ELISA format with the best performance was able to detect 10⁵ leptospires ml⁻¹ in human sera using a mAb against LipL32, the major outer membrane protein of pathogenic leptospires, as capture antibody, and a biotinylated polyclonal IgY against a pathogenic serovar of L. interrogans Icterohamorrhagiae as detection antibody. By increasing the degree of IgY biotinylation this detection limit could be improved to make the assay clinically useful.

Keywords: Leptospirosis, Leptospira, Monoclonal antibodies, Biotin, Streptavidin, Immunoglobulin Y