Development of an antibody-based capture enzyme-linked immunosorbent assay for detecting Echinostoma caproni (Trematoda) in experimentally infected rats: kinetics of coproantigen excretion

Abstract

The present study reports on the development of a coproantigen capture enzyme-linked immunosorbent assay (ELISA) for detecting Echinostoma caproni in experimentally infected rats. The capture ELISA was based on polyclonal rabbit antibodies that recognize excretory-secretory (ES) antigens. The detection limit of pure ES was 3 ng/ml in sample buffer and 60 ng/ml in fecal samples. The test was evaluated using a follow-up of 10 rats experimentally infected with 100 metacercariae of E. caproni, and the results were compared with those of other diagnostic methods such as parasitological examination and antibody titers determined by indirect ELISA. Coproantigens were detected in all the infected rats from the first day postinfection (DPI). The period of maximal coproantigen excretion was between 7 and 21 DPI. The values remained positive until 49-56 DPI, coinciding with the disappearance of the eggs in the stool samples of the infected rats. The kinetics of coproantigen detection were correlated with those of egg output. The present assay provides an alternative tool for the diagnosis of the echinostome infections. The proposed capture ELISA makes possible an earlier diagnosis than that provided by parasitological examination and indirect ELISA and also allows for the differentiation of past and current infections. Our results show that this assay can also be used to monitor the course of echinostome infections.