Development of an antigen ELISA using monoclonal antibodies against recombinant VSG for the detection of active infections of Trypanosoma evansi in animals

Abstract

Trypanosoma evansi, a haemo-flagellated protozoan parasite causes chronic wasting disease in a wide range of animals. For its diagnosis, blood smear examination is useful in clinical cases for direct identification of the parasite but in latent infection the carrier animals are difficult to screen out by conventional blood smear test. Harboring low level of parasites and showing no symptom, the carrier animals for surra can act as a source of infection. The level of parasitaemia fluctuates, especially during latent infection; moreover the antibodies which are not found early in the infection may persist even after recovery or chemotherapy. In the present study a double antibody sandwich ELISA exploring, monoclonal antibodies and hyperimmune serum, raised against recombinant variable surface glycoprotein has been developed to detect circulating trypanosome antigens. The developed antigen detection ELISA (Ag-ELISA) was evaluated using 652 blood samples collected from cattle, buffalo, equine and camel. The statistical analysis of the data showed diagnostic sensitivity and specificity at 97.4% and 96.4% respectively, with a positive-negative cut-off OD value >0.28. Furthermore, the detection limit of the assay was found to 7.15 trypanosomes per mL. The present finding revealed that the developed assay can be exploited as a potential diagnostic test in the detection of circulating trypanosome antigens and also can be used as a population screening test for multiple animal species for detection of active infection for further treatment and control of the disease.