ELISA and western blotting for the detection of Hsp70 and Hsp83 antigens of Leishmania donovani

Visceral leishmaniasis (VL) is endemic in the tropical, sub-tropical regions of Asia, Africa, the Mediterranean, Southern Europe and South and Central America. Approximately 500,000 new cases are reported annually. Classically the diagnosis of VL is confirmed by the demonstration of the parasite in aspirates of spleen, bone marrow or liver which can yield false negative results, also these methods are invasive. In this study, we aimed to evaluate the serodiagnostic potential of two heat shock proteins (Hsps) i. e. Hsp70 and Hsp83 in combination by ELISA and western blotting in mouse model. Parasite proteins were separated by SDS-PAGE and transferred to PVDF membranes which were further incubated with serum samples of infected mice for visualizing different bands. Both the bands i. e. Hsp70 and Hsp83 were simultaneously visualized in all infected groups on different post infection days. The presence of both the antigens was also detected by sandwich ELISA. The results suggest that the simultaneous occurrence of both the antigens Hsp70 and Hsp83 may be useful in serodiagnostic assay of VL as it may reduce the need for traumatic bone marrow sampling and risky spleen aspiration. © 2012 Indian Society for Parasitology.